As a prospective biomedical engineer, fluid dynamics is essential to study in order to

understand the connections between biological systems. Fluids are the main regulator within the
body; they transport nutrients to the organs, enable chemical reactions vital for proper organ
function, and facilitate muscle movement. In order to truly understand how the human body
works as the summation of small systems, it is essential to study how fluids are able to connect
these small systems in order to function as a large unit.
The goal of continuing my research project, Developing optical clearing techniques to
visualize cerebrospinal fluid flow, in Dr. Patrick Drews lab over the summer is to determine a
method to calculate the rate that fluid flows out of the brain. The role of cerebrospinal fluid
(CSF) in the brain is three-fold: it provides a layer of cushion around the brain and vertebrae, it
delivers nutrients to brain tissue and removes waste from the tissue, and it flows between the
skull and spine to account for changes in intracranial blood volume [NINHS]. CSF is formed in
the choroid plexus, located in the center of the brain. Typically, 80% of CSF secreted enters the
brains ventricular cavities [Brinker et al]. Ultimately, the CSF passes through the arachnoid villi,
which act as pathways out of the brain and into the blood [Brinker et al]. A portion of the
remaining 20% of CSF follows ventricles to the front of the brain where the fluid seeps out
through the cribriform plate, a portion of the front of the skull with tiny holes that contain the
olfactory nerves [Ethell]. While the pathways to the olfactory bulbs have been studied, it is
difficult to determine the rate at which the fluid passes through the cribriform plate and out of the
brain because we do not have a clear understanding of how CSF moves through brain tissue. By
continuing my research experience in Dr. Drews lab over the summer, I will be able to further
understand the dynamics of cerebrospinal fluid in order to determine the rate of fluid flow
through the cribriform plate.
Cerebrospinal fluid is a significant component of the bodys self-cleaning system:
according to neuroscientist Maiken Nedergaard, CSF takes out the trash in the brain during
ones sleep cycle [Nedergaard et al]. Nedergaard and her research team found that metabolic
waste products of neural activity were cleared out of the sleeping brain at a faster rate than
during the awake state [Nedergaard et al]. Proteins linked to neurodegenerative disorders like
Alzheimers are present in the interstitial space surrounding cells of the brain [Nedergaard et al].
Since neurons are highly fragile when exposed to toxic waste, the CSF recirculates through the
brain by interchanging with interstitial fluid (ISF) and flushing out interstitial proteins that cause
these neurodegenerative disorders [Nedergaard et al].
When this circulation is disrupted, the brain experiences hydrocephalus. Hydrocephalus
is a neurological disease that causes an excessive accumulation of cerebrospinal fluid within the
brain [NINDS]. When the fluid collects, the ventricles in the brain expand and generate an
immense pressure on the brain tissue [NINDS]. Understanding the mechanism and rate of CSF
flow out of the brain will allow for more effective diagnosis and treatment of hydrocephalus.
There are very few known strategies that allow researchers to examine CSF flow clearly.
To improve fluid flow visibility, I have been testing the effectiveness of various clearing
protocols, paired with dye infusions, in capturing images of CSF flow through the cribriform
plate. I have tested See Deep Brain [SeeDB], which uses water as the main solvent to limit the
use of fluorescent dyes in imaging [Ke et al]. The other protocol I have tested is a propylene
glycol-based bone clearing solution, which makes the bone transparent rather than the brain
tissue itself [Genina et al]. Figure 1 shows the comparison between clearing quality between the

SeeDB and bone clearing protocols. By observing the clarity of the olfactory bulbs, I determined
that the most effective protocols followed the SeeDB method. However, the difference in clarity
between the brain with just SeeDB and the brain with the combination of SeeDB and bone
clearing protocol are negligible. Determining the best clearing protocol will allow me to obtain
the best image of dye transport through the cribriform plate. This dye flow is representative of
the CSF flow through the plate.

Figure 1: Left to right: SeeDB, SeeDB/bone clearing

combination, bone clearing
Over the summer, I will first determine the area in the cribriform plate that CSF flows
through. While the area of the holes has been found experimentally, there are nerve bundles
present within the holes that must be accounted for. With the assistance of a grad student, I
intend to stain the nerve bundles inside the holes to determine the space they occupy. With this, I
can determine the actual cross sectional area of the CSF flow. I will then calculate the rate that
the dye flows through the cribriform plate as a function of physiological variables. Once the flow
rate function of the dye into the brain is known, CSF flow can be determined with the same
function but with different fluid constants.
With the current methodology and resources available in Dr. Drews lab, I will gain a
greater understanding of the role of fluid within the body on the micro and macro scale. This will
allow me to understand more complex features of biological fluids in the future as well as give
me a head start in my future biomedical engineering classes at Penn State. Ultimately, the
opportunity to continue my research this summer will allow me to make connections between
biological research methods and engineering research methods. I believe these connections are
necessary for me to become a world class biomedical engineer.

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Cerebrospinal Fluid Circulation." National Center for Biotechnology Information. US National
Library of Medicine, 1 May 2014. Web. 09 Mar. 2016.
<http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4016637/>.
Ethell, DW. "Disruption of Cerebrospinal Fluid Flow through the Olfactory System May
Contribute to Alzheimer's Disease Pathogenesis." National Center for Biotechnology
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<http://www.ncbi.nlm.nih.gov/pubmed/24769627>.
Genina, Elina A., Alexey N. Bashkatov, and Valery V. Tuchin. "Optical Clearing of Cranial
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09 Mar. 2016. <http://www.hindawi.com/journals/aot/2008/267867/>.
"Hydrocephalus Fact Sheet." Hydrocephalus Fact Sheet. National Institute for Neurological
Disorders and Stroke, May 2013. Web. 09 Mar. 2016.
<http://www.ninds.nih.gov/disorders/hydrocephalus/detail_hydrocephalus.htm>.
Ke, Meng-Tsen, Satoshi Fujimoto, and Takeshi Imai. "Optical Clearing Using SeeDB." BioProtocol. Bio-Protocol, 5 Feb. 2014. Web. 09 Mar. 2016. <http://www.bio-protocol.org/e1042>.
Nedergaard, Maiken, Lulu Xie, et al. Sleep Drives Metabolite Clearance from the Adult Brain
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09 Mar. 2016