CRAFT Symposium

Vi r t u a l s y m p o s i u m 2 0 2 0
Virtual Symposium
2020
Program
9 and 10 Nov 2020

Vi r t u a l S y m p o s i u m 2 0 2 0
Sponsors
• Faculty of Applied Science & Engineering, University of Toronto

• Faculty of Arts & Science, University of Toronto
• Leslie Dan Faculty of Pharmacy, University of Toronto
• Temerty Faculty of Medicine, University of Toronto
• National Science and Engineering Research Council
Sponsors
Letter from hosts

Dear symposium attendees and distinguished guests,
On behalf of the University of Toronto (UofT), the National Research Council of Canada (NRC) and the organizing
committee, we are happy to welcome you to the 2020 CRAFT Virtual Symposium.
UofT and NRC have recently established a unique and strategic partnership through the creation of a state-of-the-art
Collaboration Centre—the Centre for Research and Applications in Fluidic Technologies (CRAFT). CRAFT fosters
microfluidics-enabled device innovation for industrial and clinical applications in the areas of in vitro diagnostics,
biofabrication and organs-on-chips.
During the symposium, we look forward to exciting research presentations, lively networking, physical exercise and lots of
fun! We especially welcome our renowned keynote speakers Drs. John McDevitt (New York University), Adam Feinberg
(Carnegie Mellon University) and Valeria Orlova (Leiden University), who will share their fascinating research with us.
NRC scientists, CRAFT fellows and TOeP trainees will showcase their research in oral and poster presentations and their
entrepreneurship plans in a “business pitch” competition. Representatives from >15 companies from the entire value chain
of microfluidic technology are hosting booths in the exhibit hall.
So take advantage of these opportunities, make contacts and ask lots of questions. We want to see the chat tool humming,
lots of follow up conversations and visible engagement in CRAFT Project Award proposals. If you are an academic
researcher or clinician-scientist currently developing an exciting CRAFT Project Award proposal, this is your opportunity
to engage representatives from NRC and industry! If you are a CRAFT fellow or student, make sure you have your CV
ready and go out and mingle with industry and NRC folks, explore internship/ job opportunities and if there is a great fit
apply right away!
CRAFT is a team sport and a bottom-up initiative that has been in the making in Toronto and Boucherville for the past
15 years. We would not exist without the support of many students, faculty members, government scientists, clinicians,
representatives of technology companies, alumni and passionate administrators from academic units across UofT and its
affiliated teaching hospitals, as well as the NRC. Particular thanks to the Faculties of Applied Science & Engineering, Arts
& Science, Pharmacy and Medicine, as well as many of their departments and NRC for their continued support.
We hope you enjoy the symposium, and we look forward to seeing you next year in May 2021!
Axel Guenther Teodor Veres

Co-Director, CRAFT Co-Director, CRAFT
Associate Professor Research Director
Department of Mechanical & Industrial Engineering and BioAnalytical Micro-Nano Devices
Institute of Biomedical Engineering National Research Council of Canada
Adjunct Professor
Department of Mechanical & Industrial Engineering
University of Toronto
ii
Welcome Letter
Day 1 9 November 2020

08:30 Welcome Address / CRAFT Update Dr. Axel Guenther
• Opening Remarks Dr. Jean-Francois Houle/ Dr. Ted Sargent/

Dr. Christopher Yip
09:00 Keynote Lecture: Diagnostics Dr. Aaron Wheeler (Moderator)
Clinical decision support tools and rapid point-of-care platform Dr. John McDevitt
for determining disease severity in patients with COVID-19 New York University
09:45 Break
NRC/CRAFT Research Presentations (Session 1) Dr. Teodor Veres (Moderator)
10:00 • Multiplex droplet PCR for epigenetic-based white blood Dr. Lidija Malic
cell differential count
10:15 • Quantum dot barcode assay for multiplexed detection of Ayden Malekjahani
infectious diseases
10:30 • Advanced centrifugal microfluidic platform for the Dr. Daniel Brassard
automation of complex bioanalytical assays
10:45 • Feasibility of micro-shear wave elastography in cardiac Joseph Sebastian
microtissues
Presenters of posters 1-17 will be online

11:00 Poster Session and available to answer questions in real time
12:00 Lunch Break
NRC/CRAFT Research Presentations (Session 2) Dr. Teodor Veres (Moderator)
12:45 • Microfluidic device fabrication using biodegradable Dr. Keith Morton

polymers
13:00 • Lung airway-on-a-chip with airflow system for studying Siwan Park
epithelium and smooth muscle interactions
13:15 • Microstructured polymer substrates for bioanalytical Dr. Matthias Geissler
assays
13:30 • Development of a sample-to-answer platform designed for Alexandros Sklavounos
trauma patient monitoring based on digital microfluidics
13:45 MoveU Break MoveU Crew
14:00 Keynote Lecture: Biofabrication Dr. Axel Guenther (Moderator)
FRESH 3D bioprinting of collagen: from microfluidics to Dr. Adam Feinberg

organ biofabrication Carnegie Mellon University
14:45 Company Exhibition Hall
16:00 Awards Presentation and Closing Remarks Dr. Axel Guenther (Moderator)
iii
Program
Day 2 10 November 2020

08:30 Welcome Address Dr. Milica Radisic
08:45 Keynote Lecture: Organ-on-a-Chip Dr. Milica Radisic (Moderator)
3D Vessel-on-Chip to model vascular disease and beyond Dr. Valeria Orlova

Leiden University Medical Center
09:30 MoveU Break MoveU Crew
Presenters of posters 18-40 will be online

09:45 Poster Session and available to answer questions in real time
10:45 TOeP Business Pitch Presentations Dr. Christopher Dixon (Moderator)
10:55 • BetterMilk: animal-free dairy Noosheen Walji
11:10 • MEndR: a noval muscle endogenous repair Matthew McFee
11:25 • Cohesys: bonetape Angus Lam
11:45 Lunch Break
12:45 Invited Lecture: Biocomposites Dr. Axel Guenther (Moderator)
Transparent nanocellulose composites: a near 2D biocompatible Dr. Mohini Sain

substrate for industry 4.0 University of Toronto
13:15 Awards Presentation and Closing Remarks Dr. Milica Radisic (Moderator)
iv
Program
Abstracts
Keynote & Invited
Presentations
v
Dr. John McDevitt is a Professor in the Department of Biomaterials and Biomimetics at New York University.
He is a pioneer in the development of ‘programmable bio-nano-chip’ technologies. He has a strong track
record of translating essential bioscience, artificial intelligence and medical microdevice discoveries into real-
world clinical practice. In this capacity, he has served as the Scientific Founder for a number of diagnostic and
clinical services companies. Dr. McDevitt and his team have written more than 200 peer-reviewed scientific
manuscripts and have contributed to more than 100 patents and patent applications. This work was recognized
with the “2016 AACC Wallace H. Coulter Lectureship Award,” “Best of What’s New Award” in the Medical
Device Category by Popular Science, as well as for the “Best Scientific Advances
Award” by the Science Coalition. Dr. McDevitt’s individual honors include the
Presidential Young Investigator Award, the California Polytechnic Distinguished
Alumni Award and the Exxon Education Award.
John
McDevitt
Clinical decision support tool and rapid point-of-care platform for
determining disease severity in patients with COVID-19
SARS-CoV-2 is the virus that causes coronavirus disease (COVID-19) which has reached pandemic
levels resulting in significant morbidity and mortality affecting every inhabited continent. The large
number of patients requiring intensive care threatens to overwhelm healthcare systems globally.
Likewise, there is a compelling need for a COVID-19 disease severity test to prioritize care and
resources for patients at elevated risk of mortality. In this talk, an integrated point-of-care COVID-19
Severity Score and clinical decision support system is presented. This COVID-19 Severity Score
combines multiplex biomarker measurements and risk factors in a statistical learning algorithm to
predict mortality and is trained and evaluated using data from 160 hospitalized COVID-19 patients
from Wuhan, China. Our analysis finds that COVID-19 Severity Scores are significantly higher for
the group that died than the group that was discharged. In more recent studies, these efforts have been
expanded to include 701 patients with COVID-19 collected across practices within the Family Health
Centers network at New York University Langone Health. A two-tiered model is developed with Tier
1 using easily available, nonlaboratory data to help determine whether biomarker-based testing and/
or hospitalization is necessary. Likewise, Tier 2 predicts probability of mortality using biomarker
measurements and age. Both Tier 1 and Tier 2 models are validated using two external datasets from
hospitals in Wuhan, China comprising 160 and 375 patients, respectively. The Tier 1 and Tier 2 internal
validation had AUC (95% confidence interval) of 0.79 and 0.95, respectively; the external validation
had AUCs of 0.79 and 0.97, respectively. Collectively these promising initial models pave the way for
a point-of-care COVID-19 Severity Score system to impact patient care.
Scheduled for: 09 November 2020 from 0900 - 0945 hrs

1
Keynote Abstract
Dr. Feinberg is a Professor in the Departments of Materials Science & Engineering and Biomedical Engineering
at Carnegie Mellon University. He earned a PhD (2004) in Biomedical Engineering from the University of
Florida, where his doctoral work was focused on engineering cell-material interactions to prevent and enhance
adhesion. He was then a postdoctoral fellow at Harvard University from 2005 to 2010, where he developed
new biomaterials and cardiac tissue engineering strategies for 3-dimensional myocardial regeneration, with
a focus on stem cell-based approaches. Dr. Feinberg joined Carnegie Mellon University in the fall of 2010.
Feinberg and his team develop materials-based, regenerative strategies with the goal of engineering the self-
organization and self-assembly of various cell types into tissue structures and understanding the emergence
of higher-order function in neuromuscular and cardiovascular systems. Current efforts are focused in the
following areas: 3-dimensional nano/micro structured tissue engineering scaffolds using protein-based
materials; engineered skeletal muscle soft robotic devices and bioprosthetics; and stem cell differentiation
and engineered morphogenesis of cardiovascular tissues. Dr. Feinberg is the recipient of many distinctions,
including the NSF CAREER Award, the NIH Director’s New Innovator Award, the Ladd Research Award
from the Carnegie Institute of Technology and the Arthur Hamerschlag Career Development Professorship. He
is listed as co-inventor on 20 patents (issued/pending) and has published 45+ papers in high-profile journals
including Science, Science Advances and Nature Methods. He is also CTO and
co-founder of FluidForm, a start-up commercializing the FRESH 3D bioprinting
technology developed in his laboratory.
Adam
Feinberg
FRESH 3D bioprinting of collagen: from microfluidics to organ
biofabrication
Over the past decade, 3D bioprinting has rapidly expanded from a niche technology and into a versatile
platform for fabricating tissues with complex geometries and features ranging from the cellular to organ
length scales. Recent advances include engineering the 3D cell microenvironment, hierarchical vascular
networks in thick tissue constructs and biodegradable tissue scaffolds implanted in animal models and
human patients. However, the range of additive manufacturing technologies currently used each has
distinct advantages and disadvantages, and specifically it is critical to understand how the resolution
of these different approaches dictate structure and function of the engineered tissue constructs. Within
this context, we asked, why not build the extracellular matrix (ECM) from the bottom-up just like
cells do during embryogenesis or wound healing? To do this, we are developing new 3D bioprinting
techniques to create multiscale ECM-based structures that incorporate functional anatomy from
capillaries, to arteries, to whole hearts, while using custom-built open-source technologies. Ongoing
work is focused on developing a range of applications from a new class of ECM-based microfluidic
devices to functional cardiac tissues at the organ scale. Long-term we look to extend these approaches
to additional tissue and organ systems for in vitro disease modeling and in vivo regeneration.

2
Keynote Abstract
Dr. Valeria Orlova is an Assistant Professor in the Department of Anatomy and Embryology at the Leiden
University Medical Center (LUMC) in the Netherlands. She trained in vascular biology during her PhD and
postdoctoral fellowship at the University of Heidelberg in Germany (2003-2005) and at the Experimental
Immunology Branch at the National Institutes of Health (2005-2010) in the US. She then received a fellowship
with Drs. Christine Mummery and Peter ten Dijke at LUMC to work on differentiating vascular cells from
human pluripotent stem cells (hPSCs). The protocol she developed was patented and licensed to Ncardia
(formerly Pluriomics). She is a part of the Netherlands Organ on a Chip Initiative (NOCI) coordinated by Dr.
Mummery and an ITN European Organ on Chip (EUROoC) training network. Dr. Orlova’s current research
aims to develop realistic organ-on-chip vascular models using hPSCs and
microfluidic flow. She is on the editorial boards of Cells and Stem Cell Reports.
Valeria
Orlova
3D Vessel-on-chip to model vascular disease and beyond
Small vessel diseases are the leading cause of disability and death worldwide. The major challenge is
that they are multisystem disorders affecting different organs, such as the brain, heart and kidney. These
diseases are challenging to model in vitro because high-quality vascular cells are difficult to derive
from patients, and the local organ microenvironment which is difficult to mimic often contributes to
disease. For this reason, human induced pluripotent stem cells (hiPSCs) have become attractive sources
of patient- and organ-specific cells. We use hiPSCs to re-create blood vessels on microfluidic chips
that recapitulate micro- and macrovascular networks and the local microenvironment. We developed
efficient protocols to differentiate hiPSCs towards endothelial cells (ECs), pericytes/vSMCs and
inflammatory cells (monocytes and pro- and anti-inflammatory macrophages). We have demonstrated
that both micro- (10-50 μm) and macro-scale (250-300 μm) perfusable 3D vessels composed of
hiPSC-derived endothelial cells, pericytes/vSMCs and other non-vascular components, such as
hiPSC- derived astrocytes, can be generated inside microfluidic devices. Recently we also developed
a microphysiological system that behaves like a human “mini-heart” using cardiomyocytes, ECs and
cardiac fibroblasts all derived from hiPSCs. These mini-hearts can be produced with as little as 5000
cells and without specialized equipment. They thus represent a low-cost, low-tech platform for cardiac
drug discovery and disease modeling. Using isogenic patient hiPSC lines and 3D vessels-on-chip, we
recapitulated the phenotype of a genetic vascular disease called hereditary hemorrhagic telangiectasia
(HHT). This patient-based hiPSC model serves as proof of principle that vascular diseases can be
modeled using patient-specific hiPSCs in 3D microfluidic chips and used to identify new target cells
and possible pathways for therapy.

3
Keynote Abstract
Dr. Sain is a Professor in the Department of Mechanical & Industrial Engineering and Director of the Centre for
Biocomposites and Biomaterials Processing at the University of Toronto. He is a Fellow of the Royal Society
of Chemistry UK and the Canadian Academy of Engineers, and specializes in advanced bionanotechnology,
functional biocomposites and nature-inspired advanced devices. He holds an adjunct appointment at Lulea
University in Sweden, is an Honorary Professor at the Technical University of Bratislava in Slovakia and was
awarded an honorary degree Doctor Honoris Causa. Dr. Sain is known globally for his pioneering work on the
Biocar Initiative: in 2009, an issue of Toronto Life magazine featured his idea as second best among “25 World
Changing Ideas from the Smartest Torontonians”. He is the recipient of several awards, such as the North
American Entrepreneurship Award, the Plastic Innovation Award and the KALEV PUGI Award for innovation
and contribution to industry. He is the author of more than 600 peer-reviewed papers and is designated as a
highly-cited researcher. He holds dozens of patents and is world-renowned for his expertise in commercializing
research ideas. He has founded several spin-offs based on his research findings, including Greencore Composites
Inc., Natures Affinity Inc. and GreenNano Technologies Inc. To date, he has made
over 70 technology transfers to industry and has helped create new companies for
making a variety of products, including biomedical devices, packaging solutions,
flexible electronics and building and transportation materials.
Mohini
Sain
Transparent nanocellulose composites: a near 2D biocompatible
substrate for industry 4.0
Cellulose is a potential source of 2D-nano-material because it possesses excellent mechanical and
optical properties in its molecular state and is environmentally benign. Nanofibres derived from residual
biomass, which contains cellulose, displayed extraordinary stability under environmental stress and
bio-stability in in vivo bio-printable devices. Razor thin cellulose nanofiber composites, which are
optically active and biodegradable, are rapidly transforming biomedical and flexible device markets
due to their ability to be shaped into unique design architectures, making them functionally boundless.
The atomistic configuration changes affected by chemical, biological and mechanical attributes make
these cellulose nanofiber composites a perfect candidate for transparent, degradable nanofluidic
applications. A unique conformational architecture of transparent nanocomposite films provides glass-
like transparency and their atomic scale functional attribute as a molecular marker making them more
versatile in human health monitoring applications with targeted tracking of their mobility in a biological
system to mitigate the risk of unintended universal usage of such 2D-nanoentities, particularly in health
and food chains. Combining molecular marker and block chain tracking attributes provides unlimited
opportunities in their closed-loop life cycle in durable applications. The important functionalities of
these 2D-bionano-substrates are their transparency like glass, bendability like rubber and strength
attributes like steel. These substrates are mostly processed in aqueous systems with minimum or no
invasive procedures in healing wounds, replacing bones or delivering targeted protein. They can be
efficiently produced in a roll-to-roll process in a continuous or semi-continuous unit operations.
Scheduled for: 10 November 2020 from 1245 - 1315 hrs 4
Invited Abstract
Abstracts
Oral Presentations
5
Multiplex droplet PCR for epigenetic-based white blood cell
differential count
Lidija Malic, Jamal Daoud, Matthias Geissler, Lukic Ljuboje, Mojra Janta, Abdelrahman
Elmanzalawy, Teodor Veres
Precision Diagnostics, Medical Devices Division, National Research Council of Canada
Objectives: Perform methylation specific digital droplet PCR (ddPCR) for the purpose of white blood
cell subtyping in a multiplex format involving multiple gene targets.
Significance: Analysis of the composition of white blood cells is among the most frequently requested
laboratory tests in hematological diagnostics. Differential leukocyte count serves as an indicator for
a spectrum of diseases including infection, inflammation, anemia, and leukemia. Epigenetic markers
have recently emerged as powerful analytic tools for the study of phenotypic variations. This has led
to the investigation of cell-lineage DNA methylation patterns that correlate with human leukocyte
populations. The methods of DNA methylation analysis, however, often require costly and complex
techniques involving genome sequencing or loci-specific quantitative PCR (qPCR). Herein, a
thermoplastic elastomer (TPE) droplet microfluidic device is presented and employed for methylation
specific ddPCR. The technology has been used to delineate specific leukocyte subtypes based on cell-
lineage DNA methylation patterns with high sensitivity and specificity.
Methods: A microfluidic emulsification device fabricated from TPE promotes customizability and
cost-effectiveness of the methodology. Bisulfite-treated DNA from peripheral blood mononuclear cells
and whole blood is encapsulated in droplets with ddPCR reagents containing primers and fluorescent
hydrolysis probes specific for CpG loci correlated with WBC sub-population types. Following thermal
cycling, the droplets were imaged using fluorescence microscopy allowing accurate determination of
target copy numbers. Both qPCR and immunofluorescence (IF) staining were conducted to validate the
capacity of the ddPCR methodology to accurately determine WBC sub-populations using epigenetic
analysis of methylation sites.
Results: Highly accurate WBC subtyping has been demonstrated for frozen PBMC isolates as well as
whole blood samples in comparison with IF staining and qPCR. The ddPCR method therefore holds
the potential for clinical translation as a diagnostic tool, using blood-extracted DNA as a viable source
for differential leukocyte count. The dynamic range obtained with compartmentalization of target gene
copies was sufficient to yield ddPCR results with relatively low standard deviation and inter-variability
compared to even IF.
Oral Abstract
Quantum dot barcode assay for multiplexed detection of infectious
diseases
Ayden Malekjahani, Pranav Kadhiersan, Hannah Kozlowski, Matthew Osborne, Yuwei

Johnny Zhang, Buddhisha Udugma, Hongmin Chen, Vanessa Li, Warren C. W. Chan
Institute of Biomedical Engineering, University of Toronto, Canada
Objectives: To achieve wide spread population-level testing of infectious diseases, we require

diagnostic tools that can simultaneously detect multiple pathogens. Molecular barcoding is a technique
that can be used to create a library of unique spectral tags that identify multiple disease targets. In
this work, our objective was to develop a quantum-dot-based barcode assay and smartphone readout
device for the detection of infectious diseases (e.g. COVID-19 and other respiratory infections).
Significance: The development of a smartphone reader for quantum dot barcodes for point of care
detection would alleviate the testing burden in centralized facilities and democratize access to
diagnostic testing in low-resource settings. This is critical for the containment of diseases such as
COVID-19 and other respiratory pathogens.
Methods: Quantum-dots are semiconductor nanoparticles ideal for barcoding as they present
key advantages over conventional fluorophores with narrower emission spectra, resistance to
photobleaching and single laser excitation. Quantum dot barcodes are prepared by encoding polymeric
microbeads with variable concentrations and types of quantum dots that can be modulated to produce
spectrally distinct optical profiles. Molecules such as DNA or proteins are then functionalized to
bead surfaces that recognize genetic or proteomic material from selected pathogens. Once targets are
captured on the bead surfaces, a secondary probe attaches to the opposite side forming a sandwich
structure and confirming attachment of the target.
Results: We developed a quantum dot barcode device for detecting molecular targets of infectious
diseases. We characterized the performance of the barcodes and demonstrate the ability to excite
the barcodes using via single laser excitation and capture of the emission using the camera of a
smartphone. We further developed a software App to deconvolve the signal and to wirelessly transmit
results to a centralized database.
Oral Abstract
Advanced centrifugal microfluidic platform for the automation of
complex bioanalytical assays
Daniel Brassard, Liviu Clime, Matthias Geissler, Jamal Daoud, François Normandin,
Teodor Veres
Microfluidic Systems, Medical Devices, National Research Council of Canada
Objectives: We will present new methods to perform automated sample preparation and sample-
to-answer assays using a portable, easily-reconfigured centrifugal microfluidic platform capable of
handling diverse biological and environmental samples.
Significance: Microfluidics and lab-on-chip (LOC) technologies have been developed over the last
decade driven by the need to automate and deploy complex analytical procedures in many clinical,
industrial or research applications. LOC technologies can simplify human manipulations, reduce
associated risks of contamination, improve reproducibility, decrease reagent consumption, and enable
deployment in point-of-care settings. However, the integration of complex bioanalytical assays is a
considerable challenge in LOC, requiring not only precise control over fluid displacements but also
development of simple and reliable microfluidic devices that are compatible with low-cost mass
production.
Methods: We developed a novel centrifugal microfluidic platform that is able to control and apply
in real time air pressure pulses to microfluidics devices while they are rotating at high speed.
This combination of centrifugal forces and active pumping provides unique advantages for LOC
applications. For example, the centrifugal force enables both advanced sample preparation capabilities
(sedimentation of debris, density fractionation, etc.) and the ability to handle a wide range of sample
volumes from 1 µL to few mL with no dead volume, while the pneumatic force provides precise
control on the various liquid handling steps required by the assays (dispensing, mixing, pumping, etc.).
We will also show how the developed technology enables the integration of several complex fluidic
operations without requiring on-chip active elements or special surface treatments, opening the key
prospect to deploy complex bioanalytical assays in simple and low-cost microfluidic devices.
Results: Various automated assays developed around this technology will be discussed, ranging
from (i) isolation and purification of cellular and circulating markers (proteins, nucleic acids, etc.)
from complex matrices such as whole blood to, (ii) sample-to-answer detection of pathogens. These
demonstrations highlight the potential of this newly developed centrifugal LOC technology to deploy
a wide range of complex assays outside normal laboratory settings.
Oral Abstract
Feasibility of micro-shear wave elastography in cardiac
microtissues
Joseph A. Sebastian, Eric M. Strohm, Jérôme Baranger, Olivier Villemain, Michael C. Kolios,
Craig A. Simmons
Objectives: Cardiac microtissues, engineered from stem cell-derived cardiomyocytes, have been
developed as drug screening and testing tools. However, the widespread adoption of these models
requires the development of analytical tools that can be used to assess changes in stiffness, an
established indicator of disease and failure. We aim to develop a non-invasive and non-destructive
US technique, termed micro-shear wave elastography (μSWE), that assesses the stiffness of cardiac
microtissues in healthy and diseased states.
Significance: Current measurement techniques (e.g., atomic force microscopy, nanoindentation,

tensile testing) are invasive and incapable of monitoring microtissue stiffness over time in culture
as they are performed ex situ. μSWE would enable non-invasive, in situ stiffness measurement, thus
overcoming limitations of current microscale elasticity measurement techniques.
Methods: We developed a finite element model in COMSOL Multiphysics to predict the propagation
of US waves in 3D cardiac microtissues and various biomaterials. We used the model to evaluate US
transducers of varying central frequencies (1-10 MHz), f-numbers (1-5), and orientations (0-60° to
the vertical) to generate shear waves in various model materials (e.g., agarose, gelatin) of varying
thicknesses (1-10 mm) and cardiac microtissues. Experimentally, we have imaged biomaterials
embedded with sub-resolution scatterers (e.g., titanium dioxide) as a preliminary step toward
monitoring speckle changes due to shear wave propagation.
Results: The computational model demonstrates the dispersive effects of shear wave propagation
in simulated tissues (1 mm thickness) and the sensitivity of shear wave velocity to tissue thickness.
Moreover, the experimental results demonstrate the feasibility of a speckle tracking algorithm to
monitor shear wave propagation in these tissue-mimicking phantoms.
Scheduled for: 9 November 2020 from 1045 hrs - 1100 hrs
Oral Abstract
Microfluidic device fabrication using biodegradable polymers
Keith Morton, Matthias Geissler, Liviu Clime, Karine Turcotte, Teodor Veres
Micro and Nanofabrication, Medical Devices, National Research Council of Canada
Objectives: We present the use of biodegradable polymer materials as a sustainable alternative to

thermoplastic polymers for making single-use, microfluidic devices.
Significance: The use of microfluidic and lab-on-chip (LOC) technologies is rapidly expanding for
diverse, point-of care diagnostics. Typical implementations of POC testing for genetic or pathogenic
bioanalysis in either clinical or research settings envision wide deployment and require single-use,
disposable devices. There is therefore a strong requirement for low-cost and scalable industrial
manufacturing with injection molding using thermoplastic materials as the primary manufacturing
pathway. However, disposable microfluidic devices made from non-degrading, thermoplastic materials
create a significant, plastic waste burden. Here we present fabrication of finely microstructured,
microfluidic devices using biodegradable polymer blends of Poly Lactic Acid (PLA) and Thermoplastic
Starch (TPS)
Methods: We used standard microfabrication techniques such as photolithography and deep reactive
ion etching (Oxford) to create silicon master molds of microfluidic devices with various critical feature
sizes between 2 um and 100 um. Replica molds for hot embossing biodegradable polymer were made
by UV-curing a fluorinated resin on a glass carrier. These “working stamps” are then used to directly
hot emboss (EVG 520) biodegradable polymers.
Results: We present the fabrication of a variety of microstructures and microfluidic devices made
using PLA/TPS biodegradable polymer materials. Results include optical and SEM characterization
of microfluidic channels, reservoirs and fine-scale pillar arrays with different sizes, geometries and
aspect ratios as well as surface. We also demonstrate operation of microfluidic devices made from
biodegradable polymers including fluid handling in a centrifugal microfluidics cartridge and
continuous flow inertial focusing for particle separation.
10
Oral Abstract
Lung airway-on-a-chip with airflow system for studying
epithelium and smooth muscle interactions
Siwan Park, Edmond W.K. Young
Chronic respiratory diseases are responsible for more than 4 million deaths per year. One of the main
symptoms is airway obstruction which is a multicellular disorder caused by the abnormal airway
smooth muscle (ASM) contractions especially with the presence of inhaled particulate matters (PM).
The interactions between the airway epithelium and the ASM have been explored with experimental
animal models but the results do not translate into human airways leading to high failure rates of
the drug screening. Also, conventional bioassays have limitations in materials and geometries that
do not resemble the in vivo conditions. For the last decade, the organ-on-a-chip technologies have
been suggesting a breakthrough by enabling the human-originated cell cultures on a micro-engineered
3D environment. Studies also attempted to represent the lung airways using this technology, but the
challenge remains due to the difficulty of characterizing the airway epitheliums that vary in phenotypes
depending on the regions of the lungs. Moreover, the current PM delivery systems using transwell
inserts are limited as they lack the features of flow-driven applications like the native airways.

In order to address these issues, this study developed a micro-milled thermoplastic lung airway-on-a-
chip for coculturing the airway epithelial cells and ASM cells on a suspended hydrogel. In addition, an
airflow system that can manipulate the flow rate, humidity and temperature of the air was developed to
provide various airflows that can correlate to the aerodynamics in different regions of the lung airway.
Early results showed that after a 24-hour application of airflow on the airway epithelial cells showed
significant elongations of some motile cilia than the cells cultured in submerged or air-liquid interface
(ALI) conditions.
This study is expected to enable further investigation of the mechanobiological effects of airflow on the
lung airway epithelium. Furthermore, the flow-driven application of PM deposition using the airflow
system is expected to induce more accurate pathological responses from the interactions between the
ASM and the airway epithelium.
Scheduled for: 9 November 2020 from 1300 - 1315hrs
11
Oral Abstract
Microstructured polymer substrates for bioanalytical assays
Matthias Geissler, Lidija Malic, Keith J. Morton, Liviu Clime, Teodor Veres
Precision Diagnostics, Medical Devices, National Research Council of Canada
Objectives: This work aims at exploring the possibility of using micropillar arrays produced in a
cyclic olefin copolymer through high-fidelity microfabrication as templates for colorimetric detection
assays.
Significance: Since the production of these arrays is scalable at relatively low cost, they can provide
a suitable alternative to paper, nitrocellulose or other matrices currently used for colorimetric testing,
while facilitating integration in a polymer-based microfluidic chip.
Methods: A colorimetric DNA detection assay involving PCR-amplified gene markers for E. coli
O157 incorporating a detectable digoxigenin label, which is revealed in an immunoenzymatic process
after hybridization with target-specific oligonucleotide capture probes was implemented for proof-
of-concept demonstrations. Experimental surface chemistry was used to modify and characterize
the polymer surface so that it promotes wettability and covalent attachment of probe molecules.
Absorbance measurements and optical imaging are performed to observe and validate the formation
of color pigments during assay development. Moreover, a theoretical model is used to simulate the
changes in surface composition at the molecular level during each step of the assay.
Results: The capacity of micropillar arrays to induce wicking was used to distribute and confine
capture probes with high spatial control, making it possible to achieve a uniform signal across the
structured area. It further allowed multiple, independent probes to be arranged in close proximity
on the same substrate. Micropillar arrays also exhibit higher surface-to-volume ratios than planar
substrates, while structure-induced roughness enhances the contrast of colorimetric signal on an
otherwise transparent substrate. The relationship between microstructure and colorimetric signal was
investigated through variation of geometric parameters, such as pitch, pillar diameter, and height. Our
findings suggest that signal intensity is largely influenced by the edges of the pillars and less by their
height such that it deviates from a linear relationship when both aspect ratio and pillar density become
very high.
Scheduled for: 9 November 2020 at 1315 - 1330 hrs
12
Oral Abstract
Development of a sample-to-answer platform designed for trauma
patient monitoring based on digital microfluidics
Alexandros Sklavounos, Julian Lamanna, Dimpy Modi, M. Dean Chamberlain, Jeannie

Callum, Aaron Wheeler
Department of Chemistry, University of Toronto, Canada
Objectives: Develop a platform to determine the blood type, hematocrit and other analytes of interest
in low volume whole blood samples based on digital microfluidics (DMF). The platform was designed
to be portable with small footprint and the assays automated, reliable and with short turnaround times.
Significance: Currently, patients in need of a transfusion are initially given universal group O blood
and type AB plasma until lab results confirm their blood type leading to chronic shortage of these
blood groups. Similarly, doctors adjust the patient’s medication according to frequent tests results
performed on the patient’s blood in a lab. Consequently, there is a lot of valuable time lost until the
results reach the doctors. Therefore, bringing an instrument that could perform all these tests in near
real time to the bedside would be an extremely valuable tool for the care givers as well as help save
more lives.
Methods: For blood typing, the sample’s blood type and hematocrit are determined using
haemagglutination assays, where whole blood is mixed (using DMF) with an antibody (anti-A for
group A, anti-B for group B, and anti-D for Rhesus) specific to a certain antigen found on the red
blood cells or a chemical agglutination agent (for hematocrit) resulting in agglutination of the RBCs.
The agglutinates can be readily observed by naked eye, and the detection of agglutination can also be
automated using a digital camera and image analysis algorithms.
Results: Three novel methods were developed using digital microfluidics. The blood typing method
was calibrated on a set of 25 blood samples, and validated with a 60-sample set, showing 100%
accuracy. The method was then applied to 24 blood bank samples at the Sunnybrook hospital, operated
by a user without previous experience with microfluidics with 100% accuracy as well. For the
hematocrit determination, samples with a range of known hematocrits (20-60%) were used to generate
a calibrate curve which was then applied to a set of 12 whole blood samples from various donors.
13
Oral Abstract
Abstracts
Posters
14
Vi r t u a l s y m p o s i u m 2 0 2 0
P1: A low cost, mass manufacturable microfluidic device for
purification of small extracellular vesicles (SEVs) from whole blood
– Application to cancer liquid biopsy
Lucas Poncelet1,2, Gaetan Veilleux1, Matthew Shiu1, Daniel Sinnett2, Teodor Veres1
1
Micro & Nano-Fabrication, Medical Devices, National Research Council Canada
2
St-Justine Hospital Research Center, Montreal, Canada
Objective: Develop a microfluidic lab-on-a-chip for the automated purification of SEVs from blood
followed by cargo extraction and analysis.
Significance: Cancer represents the main cause of death due to disease in children. In the case of
childhood acute lymphoblastic leukemia (cALL), there are more than ten molecular subtypes each
with a specific prognostic risk of relapse and, in some cases, the existence of genetic actionable
alterations. The health of a patient diagnosed with cALL can decline quickly and cause death in days
to weeks without appropriate treatment. There is thus an urgent need to develop novel technologies
based on existing and new biomarkers to enable fast and accurate prognosis of cALL patients.
Methods and results: We implemented a complete immuno-magnetic capture workflow on a low-

cost polymer cartridge, the EVblade which can be mass manufactured using injection molding. The
device consists of a main fluidic processor with custom inserts enabling blood separation, filtration and
immuno-magnetic capture. Fitting the cartridge with various filters allows selection of various EVs
populations depending on the final application. The filtered plasma is then incubated with magnetic
beads functionalized with antibodies targeting a specific EV population and flowed through the
second insert, a custom on-chip magnetic capture element. After several steps of washing to remove
contaminants, the vesicles are eluted in clean buffer for downstream analysis. The purified vesicles
were characterized by TRPS and Western blotting. We observed a particle size distribution and protein
markers expected for SEVs as well as minimal contamination from other compartments.
Conclusion: We developed and characterized a low cost mass manufacturable microfluidic device for
fully automated purification of SEVs from whole blood samples. This novel device will allow us to
obtain purified SEVs preparations in a very repeatable manner from banked samples of cALL patients
as well as future patients’ samples. It will be a very useful tool to discover new biomarkers based on
SEVs cargo circulating in the peripheral blood of patients.
21
Poster Abstract
P2: WAT on a chip: a microfluidic platform for compartmental
analysis of adipose-immune crosstalk
Ayesha Aijaz1,2, Marc Jeschke1,2,3,4
1
Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada
2
Ross Tilley Burn Centre, Sunnybrook Health Sciences Centre, Toronto, Canada
3
Department of Immunology, Faculty of Medicine, University of Toronto, Canada
4
Institute of Medical Science, Faculty of Medicine, University of Toronto, Canada
Adipose tissue (AT) has been implicated as the major player in hypermetabolism following a burn
injury. During hypermetabolic state, AT undergoes a phenotypic switch energy storing white AT
(WAT) to brown AT (BAT) with increased mitochondrial mass and elevated energy expenditure.
Glucocorticoids, catecholamines and IL6 have been implicated in this switch. In addition, increased
macrophage infiltration and a shift in macrophage polarization is well established in burn-induced
browning. However, little is known about the function of regulatory T (Treg) cells as critical
determinants of post-burn hypermetabolism. Moreover, the role of AT stromal vascular fraction (where
Treg cells reside) in immunometabolic alterations is largely overlooked. Nevertheless, despite its
significance, AT pathophysiogical mechanisms in humans are still not understood, largely due to the
reliance on animal models. It is clinically impossible and unethical to conduct time-course studies on
burn patients to investigate AT dysfunction. Additionally, patient-to-patient variations create barriers to
establish clinical validation criteria. This project entails a compartmental analysis of AT morphology
and metabolism in post-burn hypermetabolism and associated alterations in resident immune cells.
We propose a WAT-on-a-chip microfluidic platform to integrate functional WAT, recapitulate its
microenvironment and induce a hypermetabolic state. In Aim 1, the microfluidic platform will
be fabricated from thermoplastics to overcome the limitations associated with protein adsorption
on polydimethylsiloxane substrates. The chip design will feature 6 tissue chambers in a parallel
configuration of 3 each for WAT microtissue formation separated from perfusable media channels by a
polyethylene terephthalate membrane. Microtissue viability, potency and quantitative analysis of key
metabolites will assessed in the effluent media. In Aim 2, WAT microtissue will be formed with/without
the stromal vascular fraction to underpin adipose-immune crosstalk. AT browning similar to post-burn
hypermetabolic state will be induced using adipose specific, 3 adrenergic agonist CL316,243. Aim
3 will target the temporal alterations in AT-resident T cells. T cell polarization to a proinflammatory
phenotype will be assessed by measuring cytokines. Changes in γδT and invariant natural killer T cells
will be evaluated. Alterations in Treg phenotype and regulation of AT inflammation will be assessed by
analyzing transcription factors, chemo/cytokines and lipid metabolism proteins. The proposed platform
provides the capability to recapitulate AT under a hypermetabolic state as a temporal signature of
readout to under pin signaling mechanisms and, the ability to perform high-throughput drug screening
and pharmacokinetic / pharmacodynamic analysis.
22
Poster Abstract
P3: High dose mesenchymal stem cells have a negative effect on
wound healing
Margarita Elloso1,2,3, Marc Jeschke1,3,4,5
1
Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada
2
Department of Laboratory Medicine and Pathobiology, University of Toronto, Canada
3
Ross Tilley Burn Centre, Sunnybrook Health Sciences Centre, Toronto, Canada
4
Department of Immunology, Faculty of Medicine, University of Toronto, Canada
5
Institute of Medical Science, Faculty of Medicine, University of Toronto, Canada
Background: The use of mesenchymal stem cells (MSCs) has been broadly studied in wound healing.
Data from different studies have shown enhanced wound healing and reduction of the inflammatory
response. The optimal dose of MSCs for wound healing is still undetermined. High doses of MSCs
may cause unexpected and unwanted results. We hypothesize that a high dose of MSCs leads to poor
quality of wound healing secondary to poor engraftment of the stem cells and apoptosis of the adjacent
cells within the wound.
Method: Using the porcine wound model multiple full thickness burns were inflicted using an
aluminum burning device. The pig was under anesthesia and analgesia throughout the procedure. The
burns were excised 48 hours post burn, to mimic the standard procedure for burn patients. During the
procedure, Integra with burn derived-MSCs (BD-MSCs) at 200,000/cm2 was applied onto the wound.
No treatment and Integra alone served as controls. Dressing change was done and wounds assessed
accordingly. Biopsies were taken to assess for cell viability, epidermal and dermal regeneration,
inflammation, vascularization and collagen density quantification.
Results: Re epithelialization was noted to be significantly slower in wounds treated with Integra
+200,000/cm2 BD-MSCs compared to acellular controls. Vancouver scar scale score was noted to be
significantly higher in wounds treated with Integra + 200,000/cm2 BD-MSCs indicating poor quality
of wound healing. (Figure 1) Cellularity was noted to be comparably higher in Integra + 200,000/cm2
BDMSCs signifying immaturity of the dermal skin tissue.
Conclusions: High dose MSCs can result in cellular overcrowding leading to apoptosis, persistent
inflammation and slower re epithelialization. This leads to an inferior quality of wound healing and
severe scarring. The ideal dose of MSCs should be determined to improve wound healing.
23
Poster Abstract
P4: Dispersing single beads into water-in-oil droplets
Jimin Guo1, Marco Serra 2, Mojra-Janta Polczynski1, Caroline Miville-Godin1, Daniel Brassard1,
Meng Zhang3, Shelby Yuan3, David Scadden4, Teodor Veres1, David Weitz3, Peter Kharchenko 2
1
Medical Devices Research Center, National Research Council Canada, Canada
2
Department of Biomedical Informatics, Harvard Medical School, USA
3
Harvard John A Paulson School of Engineering and Applied Sciences, USA
4
Department of Stem Cells and Regenerative Biology, Harvard University, USA
We designed and validated a microfluidics device to disperse single beads into a large proportion
of single droplets, enabling the capture of information from a majority of cells in droplet-based
single cell assays. Previous efforts to achieve this entailed squeezing deformable hydrogel beads
through a narrower channel with controlled pressure. This requires sophisticated pneumatic control
to simultaneously drive the flow of different phases in a set of microfluidics channels at respective
and optimal flow rates. We designed and optimized geometry and resistance of channels to achieve a
controlled flow pattern of hydrogel beads in a flow-focusing droplet generating device. This created
a close matching between frequencies of bead release and droplet generation using merely a single
pressure source, resulting in a majority of droplet to contain only one bead.
24
Poster Abstract
P5: Fetal cell isolation using laser lysis and digital microfluidics
Farhana Abbas1,2, Dean Chamberlain1,2, Julian Lamanna1,2, Michael Dryden1,2, Harrison

Edwards1,3, Aaron Wheeler1,2,3
1
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Canada
2
3
Institute for Biomedical Engineering, University of Toronto, Canada
Objectives: The goal of the project is to develop a clinically relevant process to isolate rare fetal cells
for diagnosis from maternal samples obtained noninvasively using laser cell lysis (LCL) and digital
microfluidics (DMF).
Significance: Non-Invasive methods of Prenatal Diagnosis (NIPD) provides an exciting opportunity to

monitor potential birth defects of a developing fetus without any risks. Current methods for detecting
birth defects during pregnancy are not only invasive but also increase the risk of miscarriage.
Methods: Clinical samples of cervical swabs are provided to us by our collaborators at Mount Sinai
Hospital. Samples are collected from pregnant women prior to termination of pregnancy. Maternal and
fetal cells are identified by immunofluoresence and depleted using a combination of density gradient
and immunomagentic separation. The enriched sample containing the cells of interest are allowed to
adhere onto a glass slide at specific regions ( ~3mm diameter circle). LCL, coupled with brightfield
microscopy, involves finding a fetal cell on the glass slide and focusing a short laser beam pulse in the
nanosecond range at the glass interface that the cell of interest is adhered too. The laser pulse causes
the formation of a cavitation bubble, the rapid formation and collapse of the cavitation bubble disrupts
the cell membrane and causes the release of its cellular contents. DMF is then used for the controlled
movement of small droplets that contain the contents of the lysed cell. A whole genome amplification
is then done on the lysate with the cell of interest.
Results: Most maternal cells are removed in initial enrichment steps. Of the cells that are transferred
to the DMF platform, there is about 1 fetal cell to every 50 maternal cells. Maternal cells found in the
enriched sample are mostly cervical epithelial cells and leucocytes. DNA analysis of the isolated cell
often have a maternal signature indicating maternal DNA contamination.
25
Poster Abstract
P6: Air stream agitation in microfluidic systems
Matthias Geissler, Liviu Clime, Kebin Li, Teodor Veres
Precision Diagnostics, Medical Devices, National Research Council Canada
Objectives: This work aims at exploring the possibility of using air flux to induce agitation and mixing
in confined, miniaturized systems.
Significance: Momentum transfer at the air-water interface constitutes a novel approach in

microfluidics for actively agitating liquid in low-volume cavities. The method is scalable and
compatible with parallel and multiplexed operation. The flux (and thus the strength of agitation) can
be controlled pneumatically as well as through chip design parameters.
Methods: Air flux is mediated though a pneumatic manifold connected to designated air channels
inside the microfluidic chip. Both continuous and pulsed flux are applied and their effect on flow
patterns in a liquid cavity is investigated in real-time using microscopy imaging. A theoretical model
is used for numerical simulation of momentum transfer at the air-water interface at different Reynolds
numbers.
Results: The effect of air flux-mediated agitation is investigated by following the dissolution of
fluorescent dye on a chip, showing significant reduction in the time required to reach homogenization.
The utility of the mixing approach is shown by performing nanoparticle-based capture of bacteria as
well as on-bead DNA hybridization using a dedicated microfluidic chip. Both assays provided superior
yield compared to control experiments without agitation.
26
Poster Abstract
P7: Reagentless sensing of infectious diseases through size
controlled solid-state micropores
Jenise B. Chen, Shana O. Kelley
Rapid diagnosis of infectious diseases between bacterial or viral pathogens has been crucial to prevent
misdiagnosis and antibiotic resistance. Here, I am proposing a novel technique that can provide rapid
diagnoses of infectious diseases through the combination of reagentless electrochemical detection
and solid-state micropores within a microfluidic device. Electrochemical detection can provide
rapid, quantitative signal measurements, and instrument miniaturization, while micropores can offer
a rapid size-based sorting of pathogens. Through a self-inhibited reagent depletion mechanism, the
pore size can be tuned, while creating nanostructured surfaces for electrochemical detection. These
nanostructured microelectrodes increase surface area, allowing for enhanced probe immobilization,
while increasing overall detection sensitivity. We recently developed a reagentless electrochemical
detection mechanism through an inverted molecular pendulum that is modulated by field-mediated
transport and the presence of a bound analyte. Reagentless detection of pathogens provides rapid
diagnostics without the requirement of expensive equipment or trained personnel. By developing this
reagentless micropore-based sensor, we can move towards rapid pathogen detection to help prevent
antibiotic resistance.
27
Poster Abstract
P8: Superhydrophobic thermoplastic surfaces with hierarchical
micro- nanostructures fabricated by hot-embossing
Kebin Li, J. Alejandro Hernandez-Castro, Karine Turcotte, Keith Morton, Teodor Veres
Micro-Nanofabrication, Medical Devices, National Research Council of Canada
It is not only important in fundamental research but also in practical industrial applications to control
surface wettability, for example, in the printing industry to understand the wettability of ink and toner
on various print surfaces, in self-cleaning fabrics, anti-fog windows, anti-corrosive coatings as well
as for emerging biomedical applications ranging from high-throughput cell culture platforms to liquid
flow manipulation in microfluidic devices. Much attention has recently been paid to the fabrication
of hierarchical micro- and nanostructures in polymeric materials to create superhydrophobic surfaces
with high optical transmission across the visible wavelengths for a variety of potential applications
including self-cleaning surfaces, optical components, lenses, displays, etc., and anti-wetting surfaces
against oils as well as microfluidic platforms for manipulating liquid metals.
Here we report on the fabrication of hierarchical micro-nanostructures comprising nanopillars

superimposed onto micropillars in cyclo olefin polymer (COP) materials using rapid hot-embossing
over large surface areas with single structured fields as large as 44 mm × 44 mm square. Our COP
(Zeonor 1060R) substrates are injection molded in-house in a 0.7 mm thick disc format or purchased
directly as approximately 200 μm thick ZF14-188 films. The hierarchical micro- nanostructures were
replicated in COP using an intermediate fluorinated polymer mold that facilitates the separation of
the COP substrate from the mold after hot-embossing. Moreover the intermediate mold also helps to
boost the hydrophobicity of the intrinsic COP polymer surface and hierarchical micro-nanostructures
resulting in superhydrophobic surface with a static contact angle as large as 150o. By optimizing
the overall structure design and by including additional post-fabrication fluorocarbon coating,
superhydrophobic surfaces with static contact angle as large as 167o and sliding angle as small as 2.5o
were achieved. It is possible to scale up the fabrication process to make useful application components
at low cost by using the roll to roll nanoimprinting technique. These surfaces could find potential
applications in both optical devices as well as for bioanalytical microfluidic devices.
28
Poster Abstract
P9: Molecular profiling in severe sepsis
Paul Turgeon1, Amanda Formosa1, Teodor Veres2, Claudia Dos Santos3
1
2
Medical Devices, National Research Council of Canada
3
St. Michael’s hospital, Unity Health Toronto, Canada
Epigenetic alterations are an important regulator of gene expression in health and disease. Specifically,
DNA methylation marks may provide important causal and potential biomarker information in
critically ill patients with sepsis. We have shown a total of 668 differentially methylated regions
(DMRs) corresponding to 443 genes in the blood of septic and non-septic patients. As alterations
to the DNA methylation are stable, the examination of the status of DMRs throughout the course of
sepsis can create molecular subtypes for therapeutic intervention. We anticipate that the identification
of genes and DMRs from patients at different stages of sepsis coupled with their clinical outcomes
will allow us to use molecular phenotypes to aid in determining successful clinical interventions for
patients. Peripheral blood from 22 patients has been harvested upon presentation with sepsis, upon
therapeutic intervention with ECMO (extracorporeal membrane oxygenation), and after ECMO
treatment. Globin depletion of blood samples from patients will be used for RNA sequencing.
Isolation of white blood cells from patient samples using a thermoplastic elastomer-based microfluidic
emulsification device will also be used for multiplexed, methylation-specific digital droplet PCR of
differentially expressed regions. Bioinformatic deconvolution techniques will be used to identify
molecular signatures of patient populations upon presentation and those where ECMO intervention
is beneficial. The complementary nature of RNA expression and DMRs for the blood will allow for
robust diagnostic and therapeutic targets.
29
Poster Abstract
P10: Long acting injectable and implant development utilizing
cross-linkable polyesters combined with a microfluidic approach to
establish in vitro in vivo correlation
Jack Bufton, Sungmin Jung, Zeqing Bao, James Evans, Christine Allen
Leslie Dan Faculty of Pharmacy, University of Toronto, Canada
Chronic diseases are prevalent in Canada; this is likely to increase with an ageing population. Many
chronic diseases are managed with long-term medications via repeated oral dosing. However, long-
term drug regimens put a significant economic burden on the health care system and their efficacy and
safety can be reduced, due to poor patient compliance and increased off-target effects, respectively.
Long–acting injectables and implants (LAII) offer advantages over traditional oral dosage forms
including localized drug release and minimizing dosing frequency. There is a need to expand the
number of biocompatible materials used in LAII that are biodegradable, exhibit high drug/material
ratio and sustained release.
In the first stage of this project, we produced novel cross-linked valerolactone-based implants, then
characterized their properties, including in vitro release of Paclitaxel (PTX), a model hydrophobic
drug. A pilot pharmacokinetic study in Sprague-Dawley rats following subcutaneous administration
of the implants loaded with ~13mg PTX revealed <1.5mg PTX was released by day 28, whereas the
same system released ~5.3mg under in vitro conditions.
In the second stage of this project, we are designing a release assay (i.e., SubQ-on-Chip) that exhibits
release kinetics closer to those observed in vivo to establish some degree of in vitro in vivo correlation
for LAII drug release. Such a tool would be highly useful for rapid optimization of formulations and as
quality control during manufacture. Our SubQ-on-Chip release assay, which aims to be predictive of
subcutaneous LAII drug release in vivo, will use microfluidics and a 3D cell culture. First, Fibroblasts
will be cultured then pelleted, and drug-loaded polymeric microparticles (MPs) added. Cells and
MPs will be resuspended in a cross-linking collagen matrix and transferred to a Virtoflo® chip before
collagen cross-linking is complete. Vitroflo rocking perfusion will circulate a perfusate containing
growth media and albumin through the chip. The perfusate will be sampled as appropriate to quantify
drug release from MPs over time.
30
Poster Abstract
P11: Reversible bonding of thermoplastic elastomers for cell
patterning applications
Byeong-Ui Moon, Keith Morton, Kebin Li, Caroline Miville-Godin, Teodor Veres
Precision Diagnostics, Medical Devices, National Research Council of Canada
In basic culturing techniques, cell patterning provides a simple and low-cost method for cell migration
studies. Many technologies have been developed to pattern cells within in vitro microenvironments
and study cell migration and invasion including transwell inserts, Boyden chambers and scratch
assays.
Thermoplastic materials are well known cell culture substrates. Due to their high biocompatibility,
low-cytotoxicity, minimal drugs absorption and high optical transparency, they have been used
extensively in 3D cell culture platforms, lab-on-a-chip devices, and in other biomedical applications.
Specifically, the class of thermoplastic elastomers (TPEs) have soft and flexible physical properties
and also provide advantages in the manufacturing process. TPEs are also increasingly attractive to the
researchers in the microfluidic community.
Here, we present a simple, versatile method that creates patterns using TPEs for cell migration studies.
The cell patterns are templated around a TPE feature made from a flattened TPE film using a biopsy
punch. The TPE material is strongly, but reversibly bonded to many plastic substrates, which allows us
to pattern cells in microenvironment. We examine the bonding strength of TPE on different substrates
and compare it to PDMS under various conditions. In cell migration studies, we prepare TPE posts
and position them in a 96-well plate prior to seeding fibroblast cells in the well. This simple approach
allows us to generate cell micropatterns without harsh manipulations like scratch assays, avoiding
damaging the cells. After removing the TPE posts, we examine and analyze cell migration for 2 days
in two different conditions of culture media. We also confirm that the TPE material is not toxic and
shows high viability for fibroblasts and breast cancer cells.
We demonstrate a wholly plastic-based approach to generate cell patterning without more complicated
photolithography-based processes and show its capability for cell migration study. We anticipate
this applied method can be further utilized in cell-to-cell communication studies, for example,
angiogenesis.
31
Poster Abstract
P12: Development of a defined culture medium for the maturation
of pluripotent stem cell-derived cardiomyocytes
Neal I. Callaghan1,2, Michelle Kim1, Julie Audet1, Craig A. Simmons1,2,3
1
2
Translational Biology and Engineering program, Ted Rogers Centre for Heart Research
3
Department of Mechanical & Industrial Engineering, University of Toronto, Canada
Objectives: In this study, we developed a defined culture medium to mature functional performance
of pluripotent stem cell-derived cardiomyocytes (PSC-CMs). This medium was benchmarked against
existing gold-standard media in multiple functional assays to demonstrate the advanced maturity
available in cells cultured with the new medium.
Significance: PSC-CMs hold significant promise for high-throughput drug screens and personalized
medicine. However, applications for PSC-CMs are currently limited by their immaturity in all
functional parameters including morphology, contractility, electrophysiology, Ca2+ handling, and
metabolism.
Methods: An iterative high dimensional, directed evolution algorithm was used to screen formulation
candidates for PSC-CM maturation from a factor space of over 762 billion combinations. A single
medium was chosen for intensive characterization based on high functional performance, and
compared to commercial and homemade gold-standard formulations in multiple functional parameters.
Results: Multiple iterative generations revealed a low-cost medium that induced rapid cell cycle
arrest, hypertrophy, intercalated disc formation, actin striation, and myofibril bundling in PSC-CMs to
a far greater degree than gold-standard competitor formulations. These cells were additionally further
characterized for differences in metabolic activity, proteomic profiles, and drug sensitivity.
32
Poster Abstract
P13: Polymer microfilters for cell separation applications
J. Alejandro Hernández Castro, Kebin Li, Teodor Veres
Precision Diagnostics, Medical Devies, National Research Council of Canada
Separation of cells from heterogeneous mixtures is of particular importance in the detection of

circulating tumor cells (CTCs) and filtration is a promising technique for achieving this goal in
microfluidic systems. In currently mass produced polymer microfilters (track-etched membranes),
the maximum porosity of these membranes is limited due to the random nature of the track-etching
process, which can limit the throughput of the filtration system. The development of methods allowing
low-cost fabrication of high porosity polymer membranes with customizable pore sizes is thus, of
great interest.
Here, we present a vacuum-assisted UV micro-molding (VAUM) fabrication method for making

freestanding, high-porosity polymer microfilters with controllable pore size and distribution for their
use in cell separation applications. It is a highly flexible and powerful method for fabricating low cost,
robust, large-area membranes over 9 × 9 cm2, with pore sizes from 8 to 20 μm in diameter, 20 to 100
μm in thickness, high aspect ratio (the thickness of the polymer over the diameter of the hole is up to
15: 1), high porosity, and a wide variety of geometrical characteristics. Polymeric membranes with a
hierarchical architecture made from a 200 nm-thick layer having submicron level pores (as small as
500 nm) supported by a 20 μm-thick layer has also been successfully fabricated by slightly modifying
the VAUM process. The fabricated freestanding membranes are flexible while mechanically robust
enough for post manipulation and handling, which allows them to be cut and integrated as a plastic
cartridge onto thermoplastic 3D microfluidic devices with single or multiple filtration stages.
The resulting microfilters were successfully used for the isolation of cancer cells spiked in blood
samples, as well as some preliminary tests of isolation of CTCs from cancer patients’ samples.
Nanoporous membranes (500 nm pores) have also been successfully used for the separation and
immunostaining of white blood cells (WBCs) from healthy blood samples, with the objective of using
the technology for leukemia cell trapping from blood, bone marrow, or cerebrospinal fluid (CSF)
samples in the near future.
33
Poster Abstract
P14: A high-throughput TRACER platform for screening immune
function
Nancy T. Li1, Jose L. Cadavid1,2, Alison P. McGuigan1,2
1
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Canada
2
The recent success of cancer immunotherapy has not been translated to the treatment of many solid
tumours, partially due to the complex tumour microenvironment (TME) which supports tumour
progression and reduces immune cell function. This has motivated a surge of next-generation therapies
which are armed with higher-order biological functionality. However, there is a lack of representative
and predictive models which allow for sophisticated experimentation, screening and testing phases,
which are key barriers to the development of improved cancer immunotherapies. To address this
need, we propose the development and use of a higher-throughput, 3D immuno-oncology model to
perform a functional screen of immune function modulators, thus allowing for the identification of
novel therapeutic agents or approaches for enhancing current immunotherapies. The Tissue Roll for
Analysis of Cellular Environments and Response (TRACER) model developed by the McGuigan
group is a novel, in vitro model that recapitulates the graded microenvironment within a solid tumour,
with gradients of oxygen, cytokines, and other small molecules which cannot be replicated easily in
standard 2D laboratory tools and can be adapted to fulfill this goal. The proposed research project aims
to (i) scale the existing TRACER methodology to a high-throughput platform, (ii) incorporate wild-
type and engineered immune cells to demonstrate the utility of the platform and recapitulate known
biological phenomena, (iii) perform a functional screen using an established small-molecule compound
library to identify targets with the potential to modulate immune function and tumour elimination, and
(iv) validate the functional importance of these targets. We believe that by recapitulating important
biological features of a solid tumour, we create an arena in which immune function in a tumour can
be tested, measured, and improved. We hope to contribute a novel model to this complex biological
question through the use of TRACER, expand the capability of TRACER to become a screening tool,
and further demonstrate its use as a tool for probing therapeutic efficacy of therapies. The hypothesis
underlying this study was that microenvironmental stresses imposed by solid ECM, gradients of
nutrients/metabolic waste, and direct cell-to-cell contact with diseased cell types has a direct effect on
the local behaviour of tumour-infiltrating immune cells.
Methods and Results: My current work is in designing a well-plate which generates similar gradients
to those found in the TRACER platform, and demonstrates both liquid and air-tight isolation between
individual wells. This platform allowing for individual treatment groups with (i) no diffusion of
chemical agents between treatments and (ii) no diffusion of oxygen between layers and developing a
cell-gel seeding pipeline which enables (i) parallel seeding, (ii) rapid seeding, (iii) can be automated,
(iv) user-friendly, and (v) minimizes well-to-well and batch-to-batch variation.
34
Poster Abstract
P15: Design of an islet-on-a-chip device that simultaneously
measure insulin, glucagon and GLP-1 from individual Islets
Yufeng Wang, Romario Regeenes, Jonathan V. Rocheleau
Objectives: (1) Design a microfluidic device with fluorescence sensors for insulin and glucagon
release, and with a mixing region for improvement of detection accuracy. (2) Validate the device
against standard bulk assays. (3) Measure temporal glucagon secretion relative to insulin secretion
in healthy islets. (4) Explore the impact of chronic hyperglycemia, and chronic insulin/ glucagon
stimulation on islet glucagon and insulin secretion.
Significance: Insulin, glucagon and GLP-1 are critical hormones that regulate blood glucose
homeostasis. However, the regulations of these hormones within the pancreatic islets are extremely
complex and controversial. A device that simultaneously measures real-time insulin, glucagon and
GLP-1 secretion will provide better understanding of islet biology and the pathological responses
among islet hormones in metabolic diseases such as type 2 diabetes.
Methods: (Obj 1) COMSOL modelling suggests mixing in previously developed device is inefficient.
A mixing region will be incorporated into the device and COMSOL modelling will be used to pre-
screen different device designs. With the selected design, insulin, glucagon and GLP-1 secretion
will be simultaneously monitored using fluorescence-anisotropy of a competition assay that uses
a corresponding synthetic peptide conjugated to a fluorescent dye. (Obj 2) We will test this device
against standard off-chip bulk assays including insulin and glucagon ELISA. (Obj 3) We will use our
device to simultaneously correlate the insulin, glucagon and GLP-1 responses to glucose of healthy
human islets. (Obj 4) We will use our device to simultaneously monitor insulin, glucagon and GLP-
1 secretion from human islets treated with high concentrations of glucose, or insulin/glucagon for at
least 24 hours.
Results: We are currently optimizing the design of the microfluidic device to perform the on-
chip competition assay. Preliminary COMSOL Multiphysics simulation and empirical evaluation
suggest passive mixing strategy could provide sufficient mixing while offering a simple setup of the
microfluidic system.
35
Poster Abstract
P16: High-throughput heart-on-chip platform for pharmacology
Qinghua Wu, Yimu Zhao, Naimeh Rafatian, Benjamin Fook Lun Lai, Teodor Veres,
Milica Radisic
Objectives: This work aims to develop heart-on-a-chip platforms starting from human derived iPSC-
cardiomyocytes that will enable us to understand human cardiac disease and tailor drug treatments.
Significance: We overcome manufacturing challenges to fabricate scalable and high-throughout

platform that allows in situ electrophysiological recordings from hiPSC derived cardiomyocytes
concurrently with the recording of contractile properties and recapitulate adult-like physiology for
drug testing.
Methods: A 3D printing technique was used to directly deposit microwires on electrode embedded
plate for fabricating a scalable and high-throughput platform. The soft elastic microwires was
generated to allow tissue formation and force sensors for recording tissue contraction. Conductive
carbon electrodes embedded into the multiwell plate was used to drive electrical stimulation for tissue
maturation. To demonstrate the utility of the platform, human iPSC derived cardiomyocytes were used
to form cardiac tissues in the plate device. Human iPSC derived cardiomyocytes and human cardiac
fibroblasts were mixed in a collagen hydrogel. The cell-laden hydrogel was seeded in the microwells
of the plate. Electrical field stimulation of the cardiac tissue was performed in a multiwell plate device
after cell seeding of 7 days. After tissue maturation, the effects of drug compounds on the cardiac
tissues, nifedipine and lidocaine were tested separately in the plate device for observing drug responses
on the tissues. With built-in sensors and electrodes, the contractility readouts and Ca2+ transients were
recorded according to tissue responses to the drugs.
Results: We have successfully designed new inks and developed a simple and rapid 3D printing
approach for scalable and high-throughput production of micro-wire arrayed platforms. This
manufacturing method allows us to 1) high-throughput fabrication of heart-on-a-chip devices through
rapid deposition of micro-wires in the device, 2) integration of built-in microwire sensors and
electrodes conductive for long-term electrical conditioning. A dose-dependent change in both force
and Ca2+ transients of cardiac tissues was observed in drug testing. The plate device exhibits excellent
capability to generate physiological relevance as well as high-throughput compatible systems, which
can have potential application in drug screening.
36
Poster Abstract
P17: Microfluidic Stagnation Point Devices for Analyzing Fluids
Durgesh Kavishvar, Arun Ramchandran
We encounter many fluids in our day-to-day life such as water, cooking oil, shampoo, toothpaste,
aerated drinks, ketchup, gasoline and so on. From the standpoint of flow physics, the simplest type
of the fluid is called a Newtonian fluid where the rate of deformation of the liquid is linearly related
to the applied stress. There are more complicated fluids that do not follow this proportionality and
are hence termed as non-Newtonian fluids. Multiphase materials such as mixtures of particles and
immiscible fluids are often non-Newtonian in nature. In the laboratory of complex fluids, we study
the behaviour of multiphase mixtures using different microfluidic techniques and platforms. In this
poster, I will present two microfluidic platforms that contain stagnation points in their flow fields.
A stagnation point is a point of zero velocity in the flow field, and can be used to trap soft particles
and study their hydrodynamic interactions. A single stagnation point device is termed as microfluidic
extensional flow device (MEFD). The location of the stagnation point is at the centre if the flow rates
of the liquid coming from both the inlets are equal. However, by changing the flow rates, the position
of the stagnation point can be tuned. The nature of the flow in the MEFD is such that a particle kept
at the center of the device experiences compressive stresses in the inlet direction and extensional
stresses in the outlet direction. Using this, various properties of materials can be studied. Typically,
interfacial tension, breakup characteristics, coalescence behaviours and dissolution dynamics can be
measured with this device for various emulsion systems. The platform is also suited to study other
types of soft particles such as biological cells, vesicles, elastic particles, and so on, and can work even
when the suspending medium is nearly opaque. Our laboratory has also built a two stagnation-point
device, which gives us the advantage of trapping two drops at different points. These points can be
tuned to different locations and the approach of the drops towards each other can be predefined. This
gives us the opportunity to systematically investigate head-on as well as glancing collisions between
two soft particles at prescribed approach rates, and to follow adhesion or coalescence dynamics.
Currently, we are working on a microfluidic platform that is expected to measure the yield stress of
the order of 10 - 4 Pa. As the yield stress is the measure of interactions present in the fluid, it can be
used to characterize the constituent elements of the material which contribute to the yield stress. It is
shown in the literature that blood has a low yield stress (a few mPa), and its magnitude is sensitive to
intercellular interactions. The interactions, in turn, are dependent on the concentrations of long-chain
proteins in blood, which are affected significantly by the occurrence of certain diseases. Ultimately,
this device will be developed into a point-of-care testing instrument for rapid detection of the diseases
based on the measurement of the yield stresses.
37
Poster Abstract
P18: Toward brain-on-a-chip: 3D printed layered cortical column
construct
Jianfeng Li1, Ka My Dang1, Wesley D. Sacher1, Michael G. K. Brunk1, Joyce K. S. Poon1,2
1
Department of Nanophotonics, Integration, and Neural Technology, Max Planck Institute of
Microstructure Physics, Halle, Germany
2
Department of Electrical and Computer Engineering, University of Toronto, Toronto, Canada
After more than 50 years, Moore’s Law is ending because of the physical restrictions encountered by
the semiconductor electronics. For inspiration of future computing devices and architectures, we can
look to nature’s most energy-efficient “computer”, the brain. Biological brains are more proficient at
learning and cognition compared to standard von Neumann architecture based computers. However,
physically realizing sophisticated neuromorphic constructs with well-interconnected artificial neurons
and synapses is still challenging. Here, we propose to use 3D bioprinting to build up an essential
brain computing unit, cortical column. Bioprinting enables complex human tissue architectures to be
constructed from living cells. In our planned study, we propose to 3D print layered cortical column
structure with embedded microfluidic channels to maintain inner neural cell viability and functionality.
The fabricated constructs should have well interconnected neurites, and neural circuits are formed
across discrete layers in a controlled way. The 3D printed layered cortical column can be integrated
within a chip, serving as an important computing component in the next generation artificial computing
architecture.
38
Poster Abstract
P19: Microfluidic arrays of skin spheroids for high-throughput
screening of active ingredients in cosmetic products
Zhengkun Chen1, Albert Gevorkian1, Eugenia Kumacheva1,2,3
1
Department of Chemistry, University of Toronto, Toronto, Canada
2
3
Objective: This project aims at designing and developing an organ-on-a-chip device to grow massive
arrays of skin spheroids. The successful fabrication of such a platform will then be utilized for high-
throughput screening of multiple active ingredients for skincare products.
Significance: Animal tests are intensively used in the traditional development of skincare products.
However, experiments on animal models are expensive, time-consuming, and moreover, facing ethical
criticism. In recent years, many legislative authorities around the world including Canada are working
on their regulation to ban animal tests for skincare products. Therefore, an in vitro skin model is in
great demand. Our skin spheroids-on-a-chip device is an effective and efficient solution. Combining
multicellular spheroids and microfluidic technology, fast growth of uniformly sized multicellular skin
spheroids is achieved in a microfluidic array, allowing time-, cost-, and labour efficient screening of
active ingredients. This platform can be further extended to effective screening of other substances
such as pharmaceuticals or toxins.
Methods: The massive arrays of skin spheroids were formed by the microwell-controlled formation
of droplets. The microfluidic arrays were first filled with dermal fibroblasts suspension in the hydrogel
precursor. Fluorinated oil was then pumped into the microfluidic arrays forming cell-laden droplets
inside each microwell through emulsion. After gelation of the droplets, the oil was replaced with
culture medium to allow on-chip culture of the multicellular skin spheroids. The arrays of skin
spheroids underwent a series of characterizations including live/dead staining, immunostaining on
E-cadherin, and ELISA on collagen and fibronectin synthesis.
Results: 200 uniformly sized skin spheroids were formed after 24 hours of on-chip culture. The
positive immunostaining of E-cadherin further confirmed the formation of skin spheroids as fibroblasts
aggregated and formed cell-cell junctions. Live dead staining showed good viability of multicellular
spheroids after five days of on-chip culture. Furthermore, the skin-like phenotype was observed by the
continuous production of collagen and fibronectin from the skin spheroids from Day 3 to Day 5.
39
Poster Abstract
P20: A low-cost, two-dimensional biofluid diagnostic approach:
ATR-FTIR spectroscopic microfluidics assay accessory
Tianyang Deng1, Maxime Joly2, Arthur Daignault Bouchard 2, Tyler Morhart3, Ian Burgess4,
André Bégin-Drolet2, Jesse Greener1
1
Department of Chemistry, Laval University, Canada
2
Department of Mechanical Engineering, Laval University, Canada
3
Canadian Light Source Inc., Canada
4
Department of Chemistry, University of Saskatchewan, Canada
Abstract: A microfluidics bioanalytical platform is being developed to study biofluidic and other
biological assays. This platform is a customized attenuated total reflection component serving as an
accessory for Fourier-transform infrared spectroscopy. Using motor-driven apertures, attenuated
total reflection crystal can be selectively illuminated and therefore the sample on top of which
can be selectively characterized. In addition, microfluidics devices can be built on the crystal for
sample loading. This allows multiple biofluid samples to be introduced in a parallel manner for fast
characterization, enabling high throughput in analyzing biofluid samples. Combining with deep
learning neural network and large spectra library, the platform aimed to achieve fast diagnostic of
biological assay of interest, such as determining if the concentration of COVID-19 virus is above a
medically concerning threshold. The is compatible with multiple types of spectrometers, including
portable spectrometers like ALPHA II from Bruker, allowing fast on-field diagnostic.
40
Poster Abstract
P21: Producing a confluent layer of vascular smooth muscle cells on
electrospun nanofibrous polyurethane scaffolds: Towards a tubular
cell construct
Katya D’Costa1,2, J. Paul Santerre1,2,3
1
Institute of Biomedical Engineering, University of Toronto, Ontario, Canada
2
Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research
3
Faculty of Dentistry, University of Toronto, Canada
Objective: Human primary adipose stromal cells (ASCs) differentiated into vascular smooth muscle
cells (VSMCs) were co-cultured with monocytes on polyurethane nano-fibre membranes for 2 weeks.
These cell-seeded scaffolds were assessed by immunostaining for VSMC markers - α-SMA (early-
stage), calponin (mid-stage) and smoothelin (late-stage), along with elastin and nuclear staining.
Chemical assays were used to quantify the production of extracellular matrix constituents - collagens,
elastin and glycosaminoglycans (GAGs). These scaffolds are designed to be wrapped around a
distensible silicone mandrel for mechanical conditioning, to fabricate living tubular constructs.
Significance: Patients diagnosed with atherosclerosis and type II diabetes are prone to developing
peripheral artery disease, defined as the narrowing of blood vessels in peripheral tissues. The resulting
lack of perfusion causes pain, loss of function and in severe cases, may require amputation. The
current gold standard for surgical treatment requires autologous grafts, however,
harvesting these tissues is invasive and ineffective as the patients are predisposed to compromised
vascular health. Low long-term patency rates and poor mechanical compliance of commercial grafts
stresses the urgent need for a biomimetic graft (<6mm inner diameter) for bypass surgeries.
Methods: ASCs were isolated from abdominal adipose tissue using a combination of enzymatic
digestion, centrifugation and magnetic cell-sorting followed by one week of differentiation to VSMCs
with the addition of TGF-β1 and retinoic acid. Monocytes were isolated from whole blood by density
separation. Cells were seeded on scaffolds with a surface area of 8.8 cm2 at a density of 70,000 cells/
cm2. Immunostaining was carried out with antibodies from Abcam. A hydroxyproline assay was used
to quantify collagen content, sulphated GAGs were measured with a dimethylmethylene blue assay,
and a fastin elastin assay was used for assessing elastin production.
Results: Cell-seeded scaffolds co-cultured statically, stained positive for α-SMA and calponin as well
as a weaker diffuse staining for smoothelin. Collagens, elastin and GAGs will serve as a benchmark
for further testing, following mechanical conditioning under 10% strain for three weeks, in order to
achieve a physiologically relevant compliance.
41
Poster Abstract
P22: Computational modelling of flow and drug transport in a
microfluidic device for spheroid cultures
Sina Kheiri1, Eugenia Kumacheva 2,3, Edmond W.K. Young1,3
1
Department of Mechanical & Industrial Engineering, University of Toronto, Canada
2
3
Objectives: While previous studies have investigated the effects of drug treatment on spheroids in
a tumor-on-a-chip device, none of these studies have investigated the effects of microfluidic device
geometry on drug delivery and penetration to the tumor spheroid. This paper presents a computational
model that examines the effects of geometric parameters on drug transport and tumor penetration
depth.
Significance: Here, we demonstrate a 3D computational model to study drug transport on an existing

tumor-on-a-chip device that was shown to allow high-throughput culture tumor spheroids. The model
includes advective and diffusive drug transport in different microfluidic chip geometries and sizes to
test the influence of device design on drug transport on-chip. The developed computational model also
offered insights into a full spectrum of drug transport, which eventually can be utilized for developing
strategies to optimize microfluidic chips for drug delivery and tumor treatment.
Methods: This work presents a computational model that examines the effects of geometric
parameters on drug transport and tumor penetration depth. Advection and diffusion of drug on the
chip was investigated by developing a 3D computational fluid dynamics (CFD) model in COMSOL
and performing a parametric sweep across a range of dimensions, leading to >200 simulated device
designs in ~40 h of computation time. To model drug delivery on-chip, a 3D CFD model was
developed in COMSOL Multiphysics 5.5 (Burlington,MA) by coupling two physics interfaces known
as the Laminar Flow (spf) and Transport of Diluted Species (tds) modules within the software using
time-dependent solvers.
Results: It was observed that changes in porosity impacted diffusion flux much more than advection
flux. The effects of culture chamber size on drug penetration and its mode of action were studied.
Results indicated that the size of the chamber has significant effect on the drug’s mode of transport
(advection and diffusion). Furthermore, the simulation results showed that by increasing the size of the
chamber, the major drug delivery mode changes from diffusion-dominant to advection-dominant.
42
Poster Abstract
P23: A plasmonic-assisted electrochemical platform for detection of
hydrogen peroxide released from cancer cells
Carolina del Real Mata, Roozbeh Siavash Moakhar, Imman Isaac Hosseini, Mahsa Jalali,
Sara Mahshid
Department of Biomedical Engineering, McGill University, Canada
Hydrogen peroxide (H2O2) is identified as a potential cancer biomarker due to its role in metabolic
pathways. The recently emerged electrochemical sensing can be enhanced by coupling gold
nanostructure’s plasmonic properties with the superb electrical conductivity of graphene.
Objectives: Here, we present a simple plasmonic-assisted electrochemical microfluidic device

integrating an electrode based on gold nanocavities and graphene nanosheets (NCs/Gr) for H2O2
sensing. The device was effectively used to detect released H2O2 from MCF-7 breast cancer cells with
high sensitivity.
Significance: The enhanced sensing parameters offer the possibility for implementation of this
approach towards the point of care applications. To pave the way towards industrial friendly point
of care devices, a microfluidic PDMS system, integrating a non-enzymatic, label-free, visible-light-
driven, plasmonic-assisted electrochemical gold NCs/Gr electrode is presented for detection of H2O2
released from cancer cells. This simple, portable, and sensitive approach has potential for the detection
of different types of cancer.
Methods: A simple method for electrode fabrication is proposed through a simple fabless SAM layer
of beads, metal depositions, wet-chemical etching, and exfoliated Gr drop cast. The plasmonic-assisted
electrochemical responses of gold NCs/Gr electrodes towards low H2O2 concentrations were performed
via cyclic voltammetry in a biocompatible PBS (pH 7.2) and non-enzymatic environment. Direct
detection of H2O2 and real sample experiments (MCF-7) were conducted via chronoamperometry.
Results: We reported on a plasmonic-assisted electrochemical microfluidic sensor based on a hybrid

structure of gold NCs and Gr sheets with high conductivity and reduced charge transfer resistance.
The chronoamperometry results on gold NCs/Gr showed superior properties through exceptional
electroactivity and high sensitivity towards H2O2. The sensitivity and LOD for the microfluidic chip
in biological environments were determined to be 1520 µA mM-1cm-2 in the range of 1pM-10µM and
1pM, respectively. Moreover, the assessment of MCF-7 and non-cancer cells show an increment of at
least 32% in the readings for the MCF-7 photocurrent.
43
Poster Abstract
P24: Investigating ICU-acquired weakness in a 3D human skeletal
muscle platform
Heta Lad1 , Kenneth Wu2, Judy Corea3, Tyler Saumur4, Sunita Mathur2,4, Jane Batt3,
Penney Gilbert1
1
2
Department of Physical Therapy, University of Toronto, Ontario, Canada
3
Keenan Research Center for Biomedical Science, St. Michael’s Hospital, Toronto, Canada
4
Rehabilitation Sciences Institute, University of Toronto, Toronto, Canada
Skeletal muscle is a multifunctional tissue required for several bodily functions. Despite its
remarkable regenerative capacity, certain pathologies can lead to progressive muscle weakness and
atrophy as secondary consequences; resulting in functional impairment and disability. Intensive care
unit acquired weakness (ICUAW) is a clinically detected muscle weakness in critically ill patients.
The only plausible etiology for ICUAW is the critical illness itself, often resulting in disability that is
present long after discharge, despite rehabilitation activities. It is difficult to predict which patients will
succumb to sustained ICUAW, and whether a patient will eventually recover strength. Since skeletal
muscle is a highly vascularized organ, and we are interested in how systemic factors in the blood
contribute to ICUAW by driving temporary or permanent muscle atrophy. We treated genetically
identical arrays of 3D human skeletal muscle microtissues with sera collected at a number of time-
points along the treatment of consenting critically- ill patients that may or may not develop ICUAW.
Treatment of hMMTs with sera collected within 72 hours of ICU admission with intubation and
mechanical ventilation resulted in significantly reduced hMMT myotube diameter, sarcomere integrity
and absolute passive force. Comparison of trends in hMMT form and function, when treated with
early and later timepoint serum, to clinical outcomes revealed that hMMT passive force outcomes
align with clinical outcomes of function measured by Medical Research Council sum score and motor
functional independence measure. Since blood serum is a protein rich fluid that is a primary carrier
of small molecules, the next steps will include understanding which molecules are responsible for
effects observed in culture. Successful prediction of ICUAW using blood serum aims to provide a
better understanding of mechanisms that may be involved in disease manifestation and pave the way
for new therapeutic opportunities.
44
Poster Abstract
P25: From model system to therapy – scalable production of
perfusable vascularized liver spheroids in “open-top” 384-well plate
Dawn S. Y. Lin1, Shravanthi Rajasekar1, Mandeep Kaur Marway2, Boyang Zhang1,2
1
Department of Chemical Engineering, McMaster University, Canada
2
School of Biomedical Engineering, McMaster University, Canada
Objectives: To overcome the challenge of integrating vasculature into other parenchymal tissue
models.
Significance: The incorporation of vascular networks in biological models is critical to accurately

model human diseases as well as to maintain proper tissue function in vitro.
Methods: We customized the 384-well plate (IFlowPlateTM) with a simple modification so that an
array of up to 128 perfusable vascular networks can be formed, connected, and perfused inside these
open wells. Without the physical constraints of a microfluidic channel, micro-tissue spheroids of
various sizes and quantities can be incorporated and vascularized by a microvascular bed spanning
over an area of 3.4x3.4mm.
Results: The self-assembled vessels on IFlowPlateTM had a diameter between 10-80µm, similar to
native capillary vessels. The resulting endothelial cells formed a tight vascular barrier that can confine
large fluorescent proteins (70kDa dextran) in the luminal space with minimal leakage over time. To
integrate this vascular network with larger solid tissues, we fabricated liver spheroids with a diameter
of around 200-300µm. Endothelial cells inside the spheroids were able to self-assemble within the
spheroids while the endothelial cells outside of the spheroids formed a microvascular network around
the spheroids. The vascular network around the spheroids is also perfusable, indicating that nutrients
and oxygen can be delivered with convective flow directly to the spheroids after the vasculature is
established on day 6. Finally, we demonstrated the vascularized tissues can be physically extracted
from the well plate for implantation.

To study angiogenesis, endothelial cells can be seeded inside the inlet channel against the gel
liquid interface located between the center well and the inlet channel. The extent of sprouting can
be quantified and can be used as an assay for screening pro- or anti-angiogenic drugs on this high-
throughput platform.

By growing tissues in this design, tissue spheroids can be matured in vitro and used for either high-
throughput drug screening or extracted for in vivo implantation.
45
Poster Abstract
P26: Cell invasion in digital microfluidic microgel systems
Bingyu B. Li, Erica Y. Scott, M. Dean Chamberlain, Bill T. V. Duong, Shuailong Zhang,
Susan J. Done, Aaron R. Wheeler
Microfluidic methods for studying cell invasion can be subdivided into those in which cells invade
into free space and those in which cells invade into hydrogels. The former techniques allow
straightforward extraction of subpopulations of cells for RNA sequencing, while the latter preserve
key aspects of cell interactions with the extracellular matrix (ECM). Here, we introduce “cell invasion
in digital microfluidic microgel systems” (CIMMS), which bridges the gap between them, allowing
the stratification of cells on the basis of their invasiveness into hydrogels for RNA sequencing. In
initial studies with a breast cancer model, 244 genes were found to be differentially expressed between
invading and noninvading cells, including genes correlating with ECM-remodeling, chemokine/
cytokine receptors, and G protein transducers. These results suggest that CIMMS will be a valuable
tool for probing metastasis as well as the many physiological processes that rely on invasion, such as
tissue development, repair, and protection.
46
Poster Abstract
P27: h-FIBER: Microfluidic topographical hollow fiber for studies
of glomerular filtration barrier
Ruoxiao Xie, Anastasia Korolj, Chuan Liu, Xin Song, Rick Xing Ze Lu, Boyang Zhang,
Arun Ramchandran, Qionglin Liang, Milica Radisic
Objectives: Most kidney diseases have been recognized to begin with the dysfunction of the
glomerulus, a major filtration unit of the kidney where blood is filtered to form urine. Considerable
efforts have thus been made to build an in vitro glomerulus model to better understand this filtration
unit. However, existing glomerulus-on-a-chip devices have not been able to recapitulate the
glomerular structure in the 3D configuration, which limits the physiological relevance of these models
to the native glomerulus. This study was aimed at developing an engineered 3D glomerulus model that
mimicked the structure and function of the glomerular filtration barrier.
Significance: The biomimetic 3D glomerulus model can be used to study the mechanisms of
glomerular diseases and suggest novel therapeutic paradigms. It also has the potential of being used
for drug screening and testing.
Methods: Hollow fibers with knots (h-FIBER) were produced by extruding the sodium alginate
solution (outer) and calcium chloride solution (inner) through a coaxial needle to form a luminal
structure by rapid crosslinking. The knots on the fibers were created by adjusting the flow rates to
allow the alginate solution to form a droplet at the tip of the needle before dropping into the bath
solution. Microconvex topography was incorporated into the knotted fibers via chemically induced
inflation to resemble the native configuration of glomerulus. Twenty h-FIBERs were then assembled
into a custom-made 96-well plate such that each fiber with a knot connected three wells and the knot
was situated in the middle well. Endothelial cells and podocytes were seeded within the perfusable
tubular channel (blood side) and on the knot with microconvex topography (urine side), respectively.
Results: Following long-term culture (1 month), a functional filtration barrier was established,
measured by the transfer of albumin from the blood vessel side to the urine side. Enhanced podocyte
interdigitation was demonstrated on the knot regions of h-FIBER, compared to the tube regions.
Interdigitation was further enhanced to support appropriate barrier function when the knot regions
were decorated with microtopography.
47
Poster Abstract
P28: One-step formation of protein-based tubular structures for
functional devices and tissues
Wuyang Gao1, Nima Vaezzadeh1, Kelvin Chow1, Axel Guenther1,2
1
Department of Mechanical and Industrial Engineering, University of Toronto, Canada
2
Tubular biological structures (e.g. blood and lymphatic vessels, airways, and tubules) with various
sizes, biomolecular and cellular compositions are basic functional units of all organs in animals and
humans. While low concentration, unmodified and cell-laden ECM-based biomaterials are abundantly
used in 3D cell culture, their slow fibrillogenesis kinetics and limited shear-thinning behavior have so
far limited their compatibility with extrusion bioprinting approaches. Here, the continuous, template-
free conversion of collagen, elastin, and fibrinogen solutions into tubular structures of tailored size
and radial, circumferential and axial organization is demonstrated. Importantly, we demonstrated the
formation of cell-laden collagen tubes using a biocompatible, neutral pH collagen gelation approach,
which has thus far remained a critical limitation of ECM bioprinting. The approach is enabled by a
microfluidic coaxial printhead and attached confinement for a wide range of gelation mechanisms,
including pH, temperature, light, ionic and enzymatic gelation. Both fibrillogenesis kinetics during
printing as well as stretchable, perfusible and collapsible nature of obtained tubular structures are
characterized. We demonstrated the obtained ECM tubular structures to enable unique device- and
tissue-level functional characteristics. The former is evidenced by the deterministic tube collapse and
reopening behavior, promoting in-tube flow control strategies such as all-ECM valves and pumps. The
latter is achieved by factors secreted from cells embedded within the tube wall, and endothelial or
epithelial barriers on the luminal side. We anticipate the approach to have broad applications in ECM-
based organs-on-chip and biohybrid structures, hydraulic actuators and soft machines.
48
Poster Abstract
P29: Development of an in vitro 3D microfluidic pancreatic tumor
micro environment using a machined multi-layer thermoplastic
device
Michael D. Mohan1,2, Edmond W. K. Young1,2
1
2
Pancreatic ductal adenocarcinoma (PDAC) has proven to be one of the most lethal solid malignancies
due to the heterogeneous and hostile nature of the tumor micro-environment (TME), and the highly
resistant fibrotic layer (stroma) which develops due to unregulated production of matrix proteins. The
5-year survival rate is 6% and still less than 25% after surgery. Drug development takes an average of
12 years and 1.8 billion dollars and even with immune treatments, remains challenging and expensive
due to a lack of physiologically relevant in vitro models for screening drug compounds and for patient-
specific testing. Our objective is to develop a 3D microfluidic culture platform that recapitulates
the immunosuppressive and chemoresistive nature of the TME. Here, we present preliminary data
on device design, numerical simulation, and microscale cell culture of our PDAC-on-a-chip. The
device consists of three microchannels separated by phase guides to establish surface tension-based
pinning of fluids. Minimization of phase guide height allowed maximum interaction between cells in
adjacent channels while also maintaining pinning of the gels. Devices were micro-milled over multiple
iterations, with various heights of phase guides used to test for pinning until an optimal height was
obtained. The central channel was used to load and polymerize a collagen gel that models the 3D
tumor stroma, while the side channel was coated with fibronectin and seeded with endothelial cells
for monolayer formation to serve as the vasculature of the system. COMSOL simulation data was
used to assess and predict diffusion characteristics across the monolayer boundaries and immune-
histochemistry was carried out on the monolayers to qualitatively visualize endothelial and epithelial
barrier proteins. The eventual platform will present a patient-specific in vitro drug testing system with
engineering control over essential TME characteristics.
49
Poster Abstract
P30: On-chip rotation of caenorhabditis elegans using microfluidic
vortices
Peng Pan, John D. Laver, Zhen Qin, Yuxiao Zhou, Ran Peng, Lijun Zhao, Hui Xie, John
A. Calarco, Xinyu Liu
Objectives: C. elegans is a popular model organism in biological research. However, it remains a grand
challenge to clearly and comprehensively observe all cells/neurons of C. elegans through microscopy,
even under confocal microscopy at high magnification. In this work, we would develop an easy-to-use
microfluidic device for precise rotation of C. elegans. Significance: Developed microfluidic device can
be easily used to realize precise and controllable rotation of C. elegans which could greatly facilitate
multi-perspective imaging of C. elegans and other worm species. This microfluidic-based method is
versatile as its operation is independent of the intrinsic properties of biological samples. Methods:
The layout of the microfluidic device consists of two PDMS layers of microchannels. i) The top
PDMS layer includes a microfluidic channel (green) for loading and unloading C. elegans samples,
respectively. Two torque-actuated valves are used to turn on and off the channel inlet and outlet. ii) The
bottom PDMS layer contains bifurcated microfluidic channels (blue) for regulating the generation of
microstreaming vortices inside the top microfluidic channel. When the fluid flows through the bottom
microfluidic channels, stable microfluidic vortices can be generated inside the top channel by the shear
stress from the bottom microfluidic channels. Results: By controlling the total volume flow rate of
bottom channels through a syringe pump, the worm body rotation can be controlled in both continuous
and stepwise modes. With the optimized microfluidic device, we studied the effect of syringe pump
speed ranging from 30-2500 μL/min on the worm rotation performance . The average rotation speed
increased linearly with the syringe pump infusion voltage flow rate with negligible worm drift.
Using this device, we have successfully imaged dopaminergic neurons located in the head of adult
worm (JAC882) from different orientations . By rotating the animals using our device to image from
different angles, we were able to obtain high resolution images of the neurons on all sides of the worm
at different orientations.
50
Poster Abstract
P31: A cardiac microtissue platform with integrated on-chip tissue
stiffness measurement capabilities
Christian Paniccia, Omar Mourad, Kayla Soon, Sara S. Nunes, Craig A. Simmons
Objectives: To develop and mechanically characterize a microfabricated platform that can dynamically
stretch tissue and allow for on-chip measurement of tissue stiffness.
Significance: Cardiac microtissue models hold great promise for modelling disease pathologies and
testing drugs due to their ability to recapitulate a native tissue microenvironment. For example, cardiac
fibrosis models have been developed which replicate aspects of in vivo cardiac fibrosis, including the
hallmark of tissue stiffening. However, monitoring fibrotic response in these models is limited to end
point analyses which limits understanding of disease progression. To address this need, we developed
a cardiac microtissue platform to enable non-invasive, on-chip monitoring of tissue stiffness.
Methods: A cardiac microtissue platform was designed and milled from poly (methyl methacrylate),
with a notch at one end for a polydimethylsiloxane (PDMS) anchor rod and a slot at the other end for
a free-to-move, rigid metal actuating rod. A cardiac microtissue is formed filling the well with cell-
seeded fibrin, which contract around the two rods. The microtissue platform is mounted on an actuation
platform, which includes a neodymium magnet connected to a stepper motor to laterally translate the
metal actuating rod, stretching the microtissue and causing deflection of the PDMS anchor rod, from
which the stiffness of the cardiac microtissue can be determined. Initial validation of the platform was
done by testing elastic bands (n=6) onchip, followed by testing off-chip in a tensile tester.
Results: Validation tests with elastics confirmed that stiffness measurements on-chip (10.80 ±
1.53kPa) were within 3% of those measured in the tensile tester (11.14 ± 1.33 kPa; p=0.5), indicating
that the microchip platform accurately measures the stiffness of the elastics, which was on the order of
that expected for normal cardiac microtissues. In on-going studies, microtissues composed of cardiac
fibroblasts are being monitored non-invasively for stiffness changes over time in culture. Future
studies will incorporate cardiomyocytes and TGF-β to model cardiac fibrosis.
51
Poster Abstract
P32: Development of chemically heterogenous polyurethane based
nanoparticles and peptide delivery system for chimeric peptides
Jonathan Rubianto1,2, Jonah Burke-Kleinman2,4, Michelle Bendeck 2,4, J. Paul Santerre1,2,3
1
2
3
4
Faculty of Dentistry, University of Toronto, Canada
Objectives: The goal of this study was to identify a degradable polar/hydrophobic/ionic polyurethane
(D-PHI PU) derivative that could be formed into nanoparticles and to test their compatibility with
vascular smooth muscle cells. It was hypothesized that these particles, like their thin film and scaffold
derivatives, could be used to deliver peptide therapeutics in blood contacting applications.
Significance: Atherosclerosis is one of the leading cardiovascular diseases, and interventions to date
have failed to substantially reduce the rates of re-occlusion. This is attributed to vascular inflammation
and delayed endothelial repair resulting from poor material biocompatibility, along with the limited
effectiveness of current chemotherapeutic agents.
Methods: A divinyl oligomer was synthesized with polycarbonate diol: lysine diisocyanate:
hydroxyethyl methacrylate in a molar ratio of 1:2:2. This oligomer was then mixed with methacrylic
acid and methyl methacrylate at a 1:5:15 ratio and underwent an emulsion inversion polymerization
(EIP) to generate 200 to 300 nanometer-sized particles (NPs). Metabolic activity and cell proliferation
of A10 and MOVAS cells, models of vascular smooth muscle cells, was assessed after being exposed
to different concentrations of NPs for 24 hours. Red blood cells were isolated from three donors were
exposed to the NPs to assess their hemocompatibility. The NPs were coated with peptides of interest
by taking advantage of the hydrophobic nature of the peptide. The peptide of interest is anticipated
to restrict the advance of vascular smooth muscle cells (VSMCs) and was verified utilizing a scratch
assay.
Results: Transmission electron microscopy images revealed spherical particles with a solid core.
Toxicity experiments showed that A10s and MOVAS cells do not exhibit reduced proliferation, or
changes in metabolic activity as measured by WST-1, when exposed to concentrations of 4x1010
particles/milliliter. Additionally, RBCs did not undergo hemolysis when exposed to the same
concentrations of particles. DLS and fluorescence confocal microscopy verified the coating of the
particles with peptide. The scratch assay demonstrated the same effect of slowing the advance of
MOVAS cells to the leading edge when the peptide is presented on the surface of the DPHI NPs.
52
Poster Abstract
P33: Optical growth and patterning of highly conductive silver on
ultrasmooth nanocellulose paper
Yueyue Pan, Sina Kheiri, Zhen Qin, Binbin Ying, Peng Pan, Ran Peng, Xinyu Liu
Current printed electronics with micrometer resolution requires relatively costly instruments and
sophisticated treatments. We developed a visible light assisted silver printing on nanofibrillated
cellulose paper (nanopaper), producing transparent, highly conductive silver micropatterns using a
simple experimental setup. The visible-light induced silver ion (Ag+) reduction was employed to print
highly conductive and microscale silver patterns on ultrasmooth nanocellulose paper (nanopaper). The
surface chemical group of the nanopaper ensures reliable anchoring of the silver layer, which renders
it highly conductive. We believe that this technology will find important applications in flexible
electronics and paper-based biosensing. The nanopaper was fabricated by following a previously
reported process. Then, the nanopaper was immersed into a solution of 0.15M NaNO3, 0.01M AgNO3
and 0.15M Na3Cit, and illustrated by patterned wideband visible light. The Ag printing protocol was
adapted from a previous protocol. The Ag patterns were formed through light-assisted silver nitrate
reduction into metallic silver. Because of the abundant carboxyl (−COOH) groups on the nanopaper
surface, a compact thin Ag layer can be formed on the nanopaper. The chemical composition of
the printed layer was confirmed through energy-dispersive X-ray (EDX) spectroscopy. The printed
electronics can be easily disposed by incineration. The thickness and the resistance of printed Ag strips
were measured, and the corresponding conductivity values were calculated accordingly. The highest
conductivity is 1.03×107 S/m. and the conductivity did not change significantly after 2000 times of
folding/unfolding. Functional circuits fabrications were demonstrated by a maple-leaf-shaped Ag
pattern connecting a LED in its ON/OFF state and a paper house with transparent nanopaper windows
with LED circuit. A first-order low pass filter was also built on nanopaper, and the filtering of a noisy
sinusoidal signal of 1 kHz was demonstrated.
53
Poster Abstract
P34: Generation of an arteriovenous malformation-on-a-chip
Kayla Soon1,2, Sara Vasconcelos1,2,3,4
1
Toronto General Hospital Research Institute, University Health Network, Canada
2
3
Laboratory of Medicine and Pathobiology, University of Toronto, Canada
4
Heart & Stroke/Richard Lewar Centre of Excellence, University of Toronto, Canada
Objectives: The aim of this project is to develop an in vitro microfluidic platform to model
arteriovenous malformation (AVM) for the purposes of assessing potential therapeutic targets.
Significance: There has been a lack of non-invasive treatment options for brain arteriovenous
malformations (AVM)s due to the limited understanding of its pathogenesis and the difficulty in
reproducing accurate animal models of this human disease. The KRAS mutation in endothelial cells
has been previously reported to be a predisposing factor in developing AVMs. However, the initiation
and cellular mechanisms in response to environmental factors is not well understood. Additionally,
there are no therapeutic treatments for AVMs, leaving one third of all cases untreatable. Microfluidic
devices (MFD) have been successfully used in vascular research to create functional microvascular
networks to more accurately reproduce the structural nature of a tubular interconnected vessel
network. This is largely because MFDs have the ability of creating complex customizable geometries
for highly tunable mechanical and hemodynamic properties, advantageous for mimicking the specific
microenvironment. By developing an in vitro AVM model using KRAS mutated endothelial cells, it
can take advantage of the ability to control the device parameters for a more reliable assessment of
drug therapies and improve our understanding of the AVM disease pathology and prognosis.
Methods: We are using a microfluidic design similar to Wang et al. to generate self-assembling
microvessels using a fibrin gel suspension of human umbilical vein endothelial cells (HUVEC)s and
normal human lung fibroblasts (NHLF). TRITC-dextran (10 μg/mL, 70kDa M.W) was used to test
perfusion and integrity of microvessels by introducing it to one media reservoir and observing the dye
pass through the vessel network over time. Through electroporation, a Dox inducible KRAS mutation
will be introduced into HUVECs and seeded into the MFD following the above protocol to develop
AVM-like microvessels.
Results: We found that the initial seeding, based on Wang et al. , resulted in cell aggregation at the
inlet, blocking fluid flow. After systematically seeding with lower ratios of NHLF but maintaining
the same cell density, we found that 5:1 ratio of HUVECs to NHLF developed interconnected tubular
structures without cell block as seen with high ratios. Using TRITC-dextran, the perfusability was
confirmed and there was no leakage of dye from the microvessels.
54
Poster Abstract
P35: Characterization of Electrical Cell-Substrate Impedance
Sensing (ECIS) on cell culture inserts
Alisa Ugodnikov1, Oleg Chebotarev2, Craig A. Simmons1,2
1
Institute of Biomedical Engineering, University of Toronto
2
Objective: To monitor changes in endothelial barrier integrity using electrodes in a setup compatible
with co-culture organ-on-a-chip techniques.
Significance: Barrier integrity, a key parameter for in vitro models of the blood-brain barrier, is
traditionally measured via permeability tracer assays or transendothelial electrical resistance (TEER).
These assays require invasive handling, and cannot directly assess monolayer integrity in co-culture
setups typical to organ-on-a-chip systems. As an alternative, electrical cell impedance sensing (ECIS)
evaluates monolayers directly by measuring impedance of cells grown on electrodes. ECIS has only
been implemented on solid substrates that do not allow co-culture. Here we report integration of ECIS
electrodes on porous membrane to enable non-invasive, direct monitoring of endothelial cells in a co-
culture setup, as well as validation against molecular tracer assays.
Methods: Gold electrodes were fabricated on porous membrane inserts using hot embossing and
standard UV lithography techniques. Membrane electrodes were coated with fibronectin (100 μg/mL)
to promote cell adhesion for primary human umbilical vein endothelial cells (HUVECs) and human
brain microvascular endothelial cells (HBMECs). A lock-in amplifier (SR850, Sanford Research
Systems) was used to both generate alternating current (62.5 – 64,000 Hz) and measure impedances.
Permeability was assessed using 40 kDa FITC-dextran.
Results: Frequency scanning of HUVEC monolayers (four days post-confluence on ECIS membrane
inserts) showed normalized resistance values peaking at 4 kHz, consistent with accepted 4 kHz
measurement frequency for HUVECs in conventional glass substrate ECIS setups. ECIS resistance (4
kHz) for HBMEC monolayers correlated with FITC-dextran permeability measurements, validating
the ECIS membrane method. Sensitivity was assessed by monitoring growth of HUVECs over 5 days
on working electrodes of varying sizes (d = 250, 500, 750 μm), with greatest sensitivity to changes in
barrier integrity (4 kHz) observed using the smallest electrodes.
Conclusion: Our platform produces resistance outputs and electrode size-dependent impedance trends
consistent with conventional glass substrate ECIS setups, and allows for simultaneous validation
against gold standard techniques used to evaluate co-culture systems, including molecular tracer
assays and TEER.
55
Poster Abstract
P36: Integrated ionic electronics based on horizontally-aligned
carbon nanotubes
Ran Peng, Yueyue Pan, Zhi Li, Shuailong Zhang, Aaron R. Wheeler, Shirley Tang, Xinyu Liu
The integration of natural membrane-spanning ion channel proteins or artificial nanomembrane-based

ionic diodes into planar chips to construct integrated ionic electronic circuits is very challenging.
Here, we report a new design of ionic diode, which allows chip-scale integration of iontronics (ionic
electronics), based on horizontally-aligned multi-walled carbon nanotubes (MWCNTs). Advanced
iontronic circuits including ionic logic gates, ionic rectifier and ionic bipolar junction transistor (IBJT)
created on MWCNT nanofluidic chips by stacking MWNCT ionic diodes are demonstrated.
56
Poster Abstract
P37: Investigating the role and mechanisms of extracellular vesicle
signalling in human cardiac tissue-on-a-chip models
Karl T. Wagner1,2, Milica Radisic1,2,3,4
1
2
3
Toronto General Research Institute, University Health Network, Canada
4
The Heart and Stroke/Richard Lewar Centre of Excellence, Toronto, Canada
Objectives: Ischemic heart disease and heart failure are leading causes of death worldwide. Transplant
remains a prominent treatment despite a limited supply of donor organs. To discover therapies to
treat heart disease, it is important to understand cardiac physiology at a cellular level. Recent studies
indicate that cell-signalling via extracellular vesicles (EVs) plays an integral role in the heart. It has
been suggested that EV release and uptake by cardiac cells is critical in both physiology and pathology.
The “organ-on-a-chip” industry has pioneered fabrication of human tissue models, combining cells
and biomaterials to mimic organ structure and function. These models represent a relevant, accessible
approach for investigating EVs in the heart compared to 2D cultures or in vivo studies. In this project,
the “Biowire” cardiac model will be used to investigate the role of EVs in cardiac physiology and
ischemic injury.
Significance: Adaptating the physiologically relevant Biowire model will illuminate the effects of
specific EV molecules in regulating the heart at a level not previously understood. This knowledge is
a step towards developing EV-based therapies to regenerate damaged hearts that have the potential to
revolutionize patient quality of life and reduce the societal burden of heart disease.
Methods: Biowires fabricated from differentiated induced pluripotent stem cells (iPSC) followed
by electrical maturation will be subjected to baseline functional analyses alongside compositional
profiling of tissue-secreted EVs. A model of ischemic injury will be established in Biowires, followed
by EV profile comparison to healthy tissues. EVs isolated from 2D cultures of iPSC, cardiomyocytes,
cardiac fibroblasts, endothelial cells, and healthy Biowires will be added separately to injured tissues,
assessing functional response to different sources of EVs and predicting their regenerative potential.
Results: EVs were successfully isolated and analyzed from Biowires. Media ultrafiltration decreased
background while western blotting and transmission electron microscopy confirmed the presence of
EVs in isolates. Oxygen sensors were integrated into Biowire setup to indicate physiological changes
during simulated ischemic injury for future comparison to EV profiles.
57
Poster Abstract
P38: Toward brain-on-a-chip: Microfluidic Technologies
Ka My Dang1, Jianfeng Li1, Michael G. K. Brunk1, Wesley Sacher1, Joyce K. S. Poon1,2
1
Department of Nanophotonics, Integration, and Neural Technology, Max Planck Institute of
Microstructure Physics, Halle, Germany
2
Department of Electrical and Computer Engineering, University of Toronto, Canada
We are starting a research program to develop an experimental platform for the measurement and
control of neural tissues at the microscale that integrate together electrical, optical, and microfluidic
functionalities. Microfluidic chips for neurotechnology have the advantages in flexible design,
multifunctional integration, and local micro-environment manipulation. In our proposed platform,
microfluidic channels will be used to control neuron cell growth and to implement micro-dialysis/
injection of chemicals. Microfluidic channels can be realized in silicon or in soft polymers such as
PDMS, and be simultaneously integrated with electrodes and optical waveguides for localized
multifunctional sensing and stimulation of brain tissues. The system will allow us to unveil
mechanisms in neuronal connections which will be essential in understanding activities in the brain.
58
Poster Abstract
P39: LED excitation of an on-chip imaging flow cytometer for
bead-based immunoassay
Xilong Yuan1, Todd Darcie1, J. Stewart Aitchison1, Jonathan J. D. McKendry2, Michael J.

Strain2, Martin D. Dawson2
1
Department of Electrical and Computer Engineering, University of Toronto, Canada
2
Institute of Photonics, Department of Physics, University of Strathclyde, UK
Objectives: The purpose of the project is to investigate the performance of an on-chip imaging flow
cytometry system for immunoassay and demonstrate the capability of LED excitation.
Significance: LEDs are considered an attractive alternative to laser excitation. With the development
of GaN LEDs over the last 20-30 years, LEDs are now available at wavelengths that cover from
the deep ultraviolet to the near infrared, providing flexibility in matching suitable LEDs with the
requirements of different fluorophores. Additionally, increased LED brightness can be achieved, and
LEDs can be powered by low-voltage batteries or relatively inexpensive switchable power supplies.
The proposed on-chip imaging system can detect protein biomarkers with high sensitivity and could
be potentially used for multiplex detection of antigen and antibody of infectious diseases.
Methods: A green LED is demonstrated to generate a uniform square illumination pattern for the
proposed imaging system. Aspheric lenses and filters are used to project the green light to the sample.
Bead-based fluorescent immunoassay of a sepsis biomarker (procalcitonin) is performed and detected
using the proposed system. A wide and shallow microfluidic channel is adopted for the on-chip
imaging device, allowing more bead images captured in a certain field of view. A calibration curve
of procalcitonin concentration is plotted against the fluorescent intensity using logistic regression
statistics.
Results: The LED illumination improved the uniformity of bead fluorescence intensity (coefficient of
variation is 22.4%) compared with that of a gaussian laser beam (CV is 54.7%). The limit of detection
of the proposed system for procalcitonin was determined to be 24.4 pg/mL. Future work may focus on
characterizing the multiplexing capability of the system, fabricating the device in a compact prototype
and investigating the sample preparation module of the system.
59
Poster Abstract
P40: Stretchable pressure and strain sensors based on thermoplastic
elastomer microfluidics embedded with liquid metal
Kebin Li, Karine Turcotte, Jarod Matwiy, Liviu Clime, Arron Bessoff, Matthew Shiu,
Maxence Mounier, Teodor Veres
Micro and Nanofabrication, Medical Devices, National Research Council of Canada
Abstract: Pressure and strain sensors are fundamental elements in wearable electronics for health
monitoring, human motion detection, human-machine interfaces, and soft robotics. Those sensors need
to be lightweight, flexible, bendable, stretchable and deformable to match the mechanical properties
of human skin so that they can be attached on clothing and human body for real time monitoring of
general physical activities or of kinetic motions for disease diagnosis or rehabilitation assistance.
Eutectic alloy of gallium, indium, and tin, are highly conductive, deformable and liquid at
room temperature. It has been widely applied in many applications including micro electrodes, soft
wearable robots and flexible sensors, stretchable electronics. But the flexible or stretchable substrate
used in most of those devices is based on either polydimethylsiloxane (PDMS) or Ecoflex, a type
of silicone rubber, which requires mixing of monomer with cross-link agent, followed by thermally
curing process. Moreover, the surface of the cured PDMS requires oxygen plasma treatment in order
to produce silanol terminations (SiOH) to promote the hydrophilicity of its surface, or to bond it with
another PDMS or glass substrate. Alternatively, thermoplastic elastomers (TPE) are highly stretchable
and bio-compatible, have been widely used in microfluidics in healthcare applications because of their
rapid prototyping, easy in bonding without the need of the surface treatment and low manufacturing
cost.
Here, we present the process to fabricate stretchable conductive wires based on TPE
microfluidics encapsulated with highly conductive eutectic alloy (Gallium 75.5%wt-Indium 24.5%wt
or Gallium 68.5%wt-Indium 21.5%wt-Tin10%wt). Both resistive and capacitive based pressure
and strain sensors are studied. The stretching performance of sensors was evaluated by a testing
platform in-house. Electrical resistance was measured by using four probe method. The sensitivity of
the pressure sensor depends on the mechanical properties of the TPE, thickness of the TPE layer as
well as the geometries of the sensor. A prototype of microfluidic pressure sensor with high sensitivity
that is applied for recording the arterial waveform of circulating blood, which could be used for
health monitoring will be presented. A novel, flexible and stretchable strain sensor based on TPE
microfluidics embedded with liquid metal with enhancement in both capacitance and gauge factor will
also be demonstrated.
60
Poster Abstract
Participating Companies
61
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Vi r t u a l s y m p o s i u m 2 0 2 0

9 and 10 Nov 2020

• Faculty of Applied Science & Engineering, University of Toronto

Letter from hosts

Axel Guenther Teodor Veres

Day 1 9 November 2020

• Opening Remarks Dr. Jean-Francois Houle/ Dr. Ted Sargent/

09:00 Keynote Lecture: Diagnostics Dr. Aaron Wheeler (Moderator)

NRC/CRAFT Research Presentations (Session 1) Dr. Teodor Veres (Moderator)

Presenters of posters 1-17 will be online

12:00 Lunch Break

NRC/CRAFT Research Presentations (Session 2) Dr. Teodor Veres (Moderator)

12:45 • Microfluidic device fabrication using biodegradable Dr. Keith Morton

13:45 MoveU Break MoveU Crew

14:00 Keynote Lecture: Biofabrication Dr. Axel Guenther (Moderator)

FRESH 3D bioprinting of collagen: from microfluidics to Dr. Adam Feinberg

14:45 Company Exhibition Hall

Day 2 10 November 2020

08:45 Keynote Lecture: Organ-on-a-Chip Dr. Milica Radisic (Moderator)

3D Vessel-on-Chip to model vascular disease and beyond Dr. Valeria Orlova

09:30 MoveU Break MoveU Crew

Presenters of posters 18-40 will be online

10:45 TOeP Business Pitch Presentations Dr. Christopher Dixon (Moderator)

10:55 • BetterMilk: animal-free dairy Noosheen Walji

11:10 • MEndR: a noval muscle endogenous repair Matthew McFee

11:25 • Cohesys: bonetape Angus Lam

11:45 Lunch Break

12:45 Invited Lecture: Biocomposites Dr. Axel Guenther (Moderator)

Transparent nanocellulose composites: a near 2D biocompatible Dr. Mohini Sain

Scheduled for: 09 November 2020 from 0900 - 0945 hrs

Scheduled for: 09 November 2020 from 1400 - 1445 hrs

Scheduled for: 10 November 2020 from 0845 - 0930 hrs

Scheduled for: 10 November 2020 from 1245 - 1315 hrs 4

Precision Diagnostics, Medical Devices Division, National Research Council of Canada

Scheduled for: 9 November 2020 from 1000 - 1015 hrs

Ayden Malekjahani, Pranav Kadhiersan, Hannah Kozlowski, Matthew Osborne, Yuwei

Institute of Biomedical Engineering, University of Toronto, Canada

Objectives: To achieve wide spread population-level testing of infectious diseases, we require

Scheduled for: 9 November 2020 from 1015 - 1030 hrs

Microfluidic Systems, Medical Devices, National Research Council of Canada

Scheduled for: 9 November 2020 from 1030 - 1045 hrs

Institute of Biomedical Engineering, University of Toronto, Canada

Significance: Current measurement techniques (e.g., atomic force microscopy, nanoindentation,

Scheduled for: 9 November 2020 from 1045 hrs - 1100 hrs

Micro and Nanofabrication, Medical Devices, National Research Council of Canada

Objectives: We present the use of biodegradable polymer materials as a sustainable alternative to

Scheduled for: 9 November 2020 from 1245 - 1300 hrs

Siwan Park, Edmond W.K. Young

Institute of Biomedical Engineering, University of Toronto, Canada

Scheduled for: 9 November 2020 from 1300 - 1315hrs

Precision Diagnostics, Medical Devices, National Research Council of Canada

Scheduled for: 9 November 2020 at 1315 - 1330 hrs

Alexandros Sklavounos, Julian Lamanna, Dimpy Modi, M. Dean Chamberlain, Jeannie

Department of Chemistry, University of Toronto, Canada

Scheduled for: 9 November 2020 from 1330 - 1345 hrs

Methods and results: We implemented a complete immuno-magnetic capture workflow on a low-

Scheduled for: 9 November 2020 from 1100 - 1200 hrs

Ayesha Aijaz1,2, Marc Jeschke1,2,3,4

Scheduled for: 9 November 2020 from 1100 - 1200 hrs

Margarita Elloso1,2,3, Marc Jeschke1,3,4,5

Scheduled for: 9 November 2020 from 1100 - 1200 hrs

Scheduled for: 9 November 2020 from 1100 - 1200 hrs

Farhana Abbas1,2, Dean Chamberlain1,2, Julian Lamanna1,2, Michael Dryden1,2, Harrison

Significance: Non-Invasive methods of Prenatal Diagnosis (NIPD) provides an exciting opportunity to