Cantilever

Biosensors
Mahdi Oliaee

Dr.Mohammad Rabiee
 Table of contents

What is a Market
01 Cantilever ? 02
Value

Cantilever Cantilever
03 Sensors 04 Biosensors
 01
What is a
Cantilever?
a rigid structural element that extends horizontally and
is unsupported at one end
• It can be formed as a beam, plate, truss

• When subjected to a structural load at

its far, unsupported end, the cantilever
carries the load to the support where it
applies a shear stress and a bending
moment

• Cantilever construction allows

overhanging structures without
additional support
 Applications of Cantilever Structures

Bridges: Cantilever bridges are particularly well-suited for spanning wide rivers . They are efficient and
economical solution for bridge construction.

Building Structures: often used in buildings, particularly in balconies. They allow for increased floor space .

Mechanical Devices: Cantilever structures are also employed in mechanical devices, such as cranes,
hoists, and robotic arms.

Biosensors: cantilever biosensors utilize the deflection of a microcantilever to detect

and measure specific biomolecular interactions. This technology has found
applications in medical diagnostics, environmental monitoring, and drug discovery.
02
Market
Value
13.8 billion
•
USD
The global cantilever biosensor market size expected by 2027
• Growing at a CAGR of 10.7% from 2023 to 2027
 Key Market Drivers

‫تخلخل باال‬
Increasing Demand
for POC Diagnostics
01
‫ قابل تزریق بودن‬in
Advancements
Biosensing
Rising Prevalence of
Chronic Diseases
‫تخریب پذیری‬
02 03 Technologies
 Regional Analysis
North
Asia
Europe
Pacific
America
Europe
North
Asia
America
isPacific
the second-largest
is the
a rapidly
largest
growing
marketmarket
for cantilever
for
biosensors
biosensors,
cantilever
due to
biosensors,
driven
the early
by the
driven
adoption
increasing
by the
of POC
rising
prevalence
diagnostics
demand
of and
for
chronic
thePOC
presence
diseases
diagnostics
of aand
large
inthe
emerging
number
focus on
of
economies
innovation
research and
and
in
the growing
development
focus
healthcare
on personalized
institutions. medicine
 Companies and Developers
Surface Acoustic Wave (SAW) : Piezoelectric Cantilever Biosensors:
•Abbott Laboratories •Nanonics Imaging
•Roche •Bio-MEMS
•Danaher Corporation •NanoSensors
•Siemens Healthcare •NanoPlus Technologies

Companies that sell platforms: Optical Cantilever Biosensors:

•Bruker Corporation •Biacore International
•Hitachi High-Technologies Corporation •Biosensing Technologies GmbH
•Shimadzu Corporation •Cantilever Designs
•JEOL Ltd. •Cantion B.V.
 Patent Document Over Time

NEMS Biosensors MEMS Biosensors Cantilever biosensor

 Prices
The exact price of a cantilever biosensor will vary depending on the specific features and
specifications of the device. However, as a general guideline, here is a price range for
different types of cantilever biosensors:

Microcantilever biosensors(MEMS) $100 to $10,000 per device

Nanocantilever biosensors(NEMS) $10,000 to $100,000 per device
Microarray-based cantilever biosensor systems $10,000 to $100,000,000 per system
03
Cantilever
Sensors
• Cantilever sensors respond to changes in their environment or changes on their
surface with a mechanical bending in the order of nanometers which can be detected.
• Cantilever sensors emerged from atomic force microscopy (AFM)
• AFMs can image surfaces, nanosystems or single molecules
with Angstrom resolution, manipulate molecules,
or measure forces between individual molecules.
Microfabricated cantilevers are made of silicon or silicon nitride using basically the same
micromachining methods as for computer chips .Their dimensions are in the micrometer range

• How to describe ?
Hooke’s law: 𝐹 = −𝑘𝑠𝑝𝑟𝑖𝑛𝑔 ∆𝑧

• What is 𝒌𝒔𝒑𝒓𝒊𝒏𝒈 ?

𝐸𝑤𝑡 3
𝑘𝑠𝑝𝑟𝑖𝑛𝑔 =
4𝐿3
E : Young’s modulus
Experiments with complex biological samples are W : Cantilever Width
T : Thickness
nowadays performed with cantilever arrays of up to L : Length
eight cantilevers in parallel. All are physically
identical and only differ in their surface coating.
• Scanning electron micrograph (SEM) of a
part of a microfabricated cantilever Array.
• The silicon cantilevers are 1 𝜇m thick , 100
𝜇m wide and 500 𝜇m long. They have a
pitch of 250 𝜇m.
• Cantilever beams are not directly fixed to the
bulk silicon,but attached via a thicker bar to
a solid platform
• Such a geometry facilitates cantilever
functionalization and prevents spreading of
functionalization liquids from one cantilever
to the other
 Okey, what about readout of the Cantilever
bending ?!
•Optical readout :
a) uses a laser that is reflected from the tip of the cantilever
b) detected by a position-sensitive photodetector
c) suffers from drawbacks such as artifacts due to changes in the refraction index of the medium
•Diffractive optical element:
a) uses a holographically patterned thin photopolymer layer that is placed on a polymer layer, forming
a bi-layer cantilever
b) The photopolymer layer acts as a diffractive optical element that diffracts light at different angles
depending on the cantilever deflection, The diffraction efficiency of the element is measured by a
photodetector
c) low-cost readout, and can achieve high sensitivity
•Piezoresistive:
a) uses a thin metal or semiconductor layer on the cantilever and acts as a strain gauge
b) resistance of the layer changes with the cantilever deflection due to the piezoresistive effect
c) The resistance change is measured by an electrical circuit
d) enables simple measurements on non-transparent liquids , such as blood
e) can be integrated with microfluidic systems
•Capacitive:
a) uses a pair of electrodes that are attached to the cantilever and a fixed substrate, forming a variable
capacitor
b) The capacitance changes with the cantilever deflection due to the change in the gap between
the electrodes , change is measured by an electrical circuit
c) high sensitivity and low power consumption but requires precise alignment
• To improve readout cantilevers can be covered with a thin reflecting metal layer
(typically gold) on one side as a substrate.
• The gold coating is normally applied directly before the functionalization of a cantilever to
avoid contaminations by a longer exposure to air
• Thereby, a nanometer-thin adhesion layer of Ti or Cr
ensures a proper adhesion of the several 10 nm thick
gold layer to the cantilever
• In the Dynamic or Resonance mode, cantilevers are
excited close to their resonance frequency , When
an additional mass is attached to the oscillating
cantilever, its resonance frequency changes

Mass Resonance frequency

In Resonance mode :
1 𝑘𝑠𝑝𝑟𝑖𝑛𝑔
𝑓=
2𝜋 𝑚∗
With optimized geometries and under
Effective mass : Cantilever
ultra-high vacuum one can measure mass
geometry and mass distribution
changes in the resonant mode down to the
single molecule level.

For biomolecules: viscous damping of the

oscillation decreases mass resolution
In Heat mode :

Cantilevers which are coated by a thin metal

layer can also act as bimetallic sensors

bend 100 nm for a 1 K temperature difference

when covered with some 10 nm of gold.

With optimized setups, temperature changes

smaller than 10−5 K can be detected and
calorimetric measurements with a sensitivity
of 10−5 J are possible
Surface stress or Static mode:

by far the most common mode for biosensing experiments

changes in the environment around the Cantilever create a mechanical stress in the surface
leads to an expansion or contraction of the cantilever surface

If this stress acts only on one side of the cantilever, then the asymmetrically stressed structure
will bend and the cantilever will deflect.
when talking about surface stresses detected by cantilever sensors we always refers to a
change in surface stress, but not to an absolute stress
Surface stress or Static mode:

The most commonly used formula to relate cantilever deflections to surface stresses is
Stoney’s formula
Elasticity modulus thickness
𝐸𝑡 2
∆𝜎 = ∆𝑧 Detected cantilever deflection
3 1 − 𝜈 𝐿2
Difference in surface stress
Poisson ratio Effective length

Length Deflection Thermal Oscillation amplitude

Tickness
∝ Sensitivity
BUT The noise in Detection
 04
Cantilever
Biosensors
 Cantilever Biosensors Deflection
• increasing the surface charge on one side of a cantilever
• changes of pH followed by a protonation or deprotonation of the surface which lead to an
electrostatic repulsion of surface groups which leads to an expansion of a cantilever
surface A: immobilized DNA changed from : flexible, single stranded
to stiff, hybridized, double stranded which Reduces lateral
steric interactions ( Repulsive ) between DNA
B: deprotonation of molecular groups on the cantilever
surface (caused by change in pH) creates negative surface
charges and an electrostatic repulsion
 DNA hybridization process

Alkanethiol-based self-assembled
monolayers on a gold-coated
cantilever platform
 Sensor surface Functionalization
Physical Adsorption:
• Antibodies adsorption onto gold surfaces using electrostatic interactions.
• DNA strands adsorption onto silica surfaces using hydrogen bonds.
• Proteins adsorption onto surfaces using van der Waals forces.
Covalent Binding:
• Thiol-functionalized biomolecules adsorption onto gold surfaces using thiol chemistry.
Self-Assembly:
• Peptides can self-assemble into nanoarrays on gold surfaces.
• DNA can self-assemble into structures on surfaces with specific sequences.
• Lipids can self-assemble into bilayers on surfaces.
Sensor surface Functionalization
Coating or functionalizing a sensor is a critical preparation step because the recognition layer
will define the application and performance of a sensor.

Challenges:
Making the coatings such that the biomolecules are tightly attached to the sensor surface but
are still flexible and functional as in their natural environment

In surface stress mode Binding target molecules only on one side

One surface should provide the receptors for the target molecules, whereas the
other should prevent any specific or unspecific adsorption of sample molecules

Perfect Coating should be robust against changes in buffer and temperature and
ideally withstand repetitive detection and cleaning cycles.

Protein resistive monolayer coating

Gold surface

Silicon oxide surface

Receptor molecules

Ligands
Sulfur groups have high affinity(high tendency to bind) for gold, so gold surface
can be modified by thiolabeled nucleic acids or proteins exposing cysteines at
their surface .
On the other hand, thiolated poly(ethylene glycol)s can act as inert layers,
preventing molecular adsorption

In addition, highly positively charged molecules can be electrostatically bound

to the negatively charged silicon dioxide

 How to expose Cantilevers to
specific biomolecules?!
• Be incubated in individual glass capillaries or standard pipette tips filled with coating
molecules
• Be dipped in the channels of an open microfluidic network
• Spotted with microliter drops of receptor molecules using an ink jet type of device

Hinge region Special care for homogeneous coating

 MEMS and NEMS
MEMS (Micro Electromechanical Systems) and NEMS (Nano Electromechanical Systems) are
a cantilever sensors with remarkable sensitivity to the slightest changes in their environment

Material Selection for MEMS and NEMS

silicon nitride (𝑆𝑖3 𝑁4 ), and gold are commonly used materials for MEMS cantilevers due to their
excellent mechanical stability, low mass, and resistance to corrosion.

For NEMS cantilevers, materials like silicon monoxide (SiO), single-walled carbon nanotubes
(SWCNTs), and graphene are preferred due to their exceptional mechanical properties and ability
to achieve ultra-high sensitivity.
 What’s the difference ?

NEMS cantilever sensors offer the highest sensitivity and resolution, but they are
also the most challenging to fabricate and operate.

MEMS cantilever sensors are a more practical choice for many applications, as they
offer a good balance of sensitivity, range, and reliability
 Feature NEMS MEMS

Size Nanoscale Microscale

Sensitivity Ultra high High

Dynamic Range Narrow Wider

Response time Fast Medium

Noise Level Low Medium

Power requirement Low Medium

Fabrication Electron beam Lithography, Photolithography,

Technique FIB milling Deposition

Dynamic range refers to the ratio of the largest and smallest signals that the sensor can
accurately measure
Why NEMS has better Response time and
noise level ?
Nanoscale dimensions reduce their thermal noise as the smaller structures
Smaller Dimensions
dissipate heat more efficiently

Materials used for NEMS has higher elastic modul this high stiffness
High Elastic Modulus
minimizes the deflection of the cantilever due to environmental vibrations

NEMS cantilevers are typically designed with low damping properties enables the
Low Damping
cantilever to respond quickly to changes

The Q factor of a resonant system describes its ability to store energy and
High Q Factor
release it smoothly NEMS cantilevers can maintain their resonant frequency with
minimal damping better than MEMS
 Model Experiment
After a cantilever array has been cleaned, covered with gold and functionalized

the array is transferred to a liquid cell with an inlet and an outlet and a volume of typically less than 100 𝜇𝐿.

Buffer is injected either by hand or via an automated delivery system with pumps and valves

Readout lasers are aligned onto the position-sensitive detector

Temperature control is an advantage due to silicon–gold bimetallic effects.

 What is Cantilever Drift ?
Drift refers to the long-term changes or instability in the sensor’s output when the input is constant.
Since the cantilevers have so far been subjected to several changes from ambient to liquid
environments due to cleaning or functionalization they are most likely not in an equilibrium state
and will show a strong drift of approximately several nanometers per minute.

How much Drift is acceptable ?

an experiment is started when the drift reduces to about a few nanometers per hour
 Deflection versus time graph

Functionalized with receptor 1. Injection of negative control

2. Injection of sample molecules
Reference Cantilever
3. Same injection to verify saturation and completion of reaction
4. Buffer injection leading to unbinding
 Cantilever biosensor applications
• The monitoring of digestion and ligation of DNA on surfaces and the activity of a polymerase enzyme
by cantilever sensors
• Proteins sensor for prostate specific antigen or PSA
• for detecting glucose using glucose oxidase enzymes
• for cardiac biomarkers (which are proteins indicating a heart attack)
• pH and Temperature Changes.
• Food Analysis which is for the detection of contaminants and quality control.
• DNA Hybridization.
• Antibody-Antigen Interactions.
• Adsorption of Bacteria.
 Cantilever biosensor applications
The first applications of The development of
Single cantilever sensors NEMS/MEMS cantilever-
for biological systems based biosensors

1994 1996 2000 2008 Now!

The development of first clinical diagnostics, food analysis,

Microcantilever-based biosensing experiments bioprocess, and environmental
Biosensors with cantilever arrays monitoring
The first applications of cantilever sensors for biological systems were reported in 1996 with
single cantilevers H.-J. Butt, J. Colloid Interface Sci., 1996, 180, 251–260

Lockheed Martin Energy Systems Inc

US5719324A
 The Road ahead …
• Advances can be expected in transducing stress into a larger cantilever deflection or improving
surface coatings so that even lower molecular concentrations create enough stress.

• Cantilever array biosensors sensitivity is comparable with existing label-free technologies. They now
have to prove that their every-day performance and ease of handling can also compete with
established methods outside the specialized labs where they have been developed
Conclusions
and
Outlook
Cantilever sensors have already shown an impressive performance
Their major advantages are their:

• wide field of application

• they are label-free
• can be microfabricated
• need only small sample volumes for their operation
• Their response is comparable with other label-free biosensing methods
• general setup can be easily modified to detect a variety of different parameters
 So What’s the Problem ?!
Problems which still delay a general use of cantilever sensors are:
in particular, their need for a sophisticated surface functionalization and the lack of a
theoretical description.
The fact is that in cantilever sensor research, the theoretical description is at the
moment lagging behind the technical developments and the progress in applications
especially for the surface stress mode.

But these are not fundamental problems though !

And already being addressed by researchers
When ignoring all the technical
details , the simple but
fascinating fact remains that
thin layers of soft molecules are
able to bend solid state devices.
References
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5 ) C. Ziegler, Anal. Bioanal. Chem., 2004, 379, 946–959.
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Thank You
for Your
Attention!