A Novel Microfluidic PCR
Platform for Rapid Tissue Typing
Immunofluidics
NAME
Brandon
Wong
Philip
Chao
Irfanali
S.
Kermalli
Jay Le
Pere
Daniel
Lorey
UCI SCHOOL/
OTHER
AFFILIATION
Henry Samueli
School of
Engineering
Henry Samueli
School of
Engineering
Henry Samueli
School of
Engineering
Henry Samueli
School of
Engineering
Henry Samueli
School of
Engineering
GRADUATE/UNDER- MAJOR/MINOR
GRADUATE
FIELD OF STUDY
EMAIL ID
Undergraduate
BME
brandogw@uci.edu
Undergraduate
BME/Materials
Minor
pchao@uci.edu
Undergraduate
BME/Management
Minor
ikermall@uci.edu
Undergraduate
BME
jlepere@uci.edu
Undergraduate
BME/Materials
Minor
dlorey@uci.edu
Division: Diagnostic Devices/Methods
Page 1
Abstract
The ImmunoFluidics Chip is a device that leverages cutting edge diagnostic technologies to
assist doctors in finding a donor-recipient match during organ transplantations. Tissue matching
has been proven to make a significant difference on survival rate in organ transplant recipients.
However, due to the urgent nature of these transplantations, there is not enough time for tissue
typing to be done. Consequently, surgeons choose to put a healthy organ into a patient rather
than taking the time to match the organ, potentially lowering its viability. The ImmunoFluidics
Chip leverages microfluidic technology to be able to perform rapid tissue typing. By performing
tissue typing faster, more information about the organ can be determined, letting surgeons
make a better decision before transplantation of the organ.
Background and Clinical Need
Every year, organ transplantation saves the lives of thousands of patients. In 2009, records
show that roughly 30 thousand organ transplantations were performed in the United States
alone1. Even though this procedure has been prevalent in clinical practice for many decades,
there are still areas of improvement that can result in higher patient survival rate2. Specifically,
tissue typing has been proven to make a significant difference on reducing the amount of
immunosuppressant drugs used, shortening the recovery time after transplantation, and
increasing the probability of survival post-implantation2. However, due to the urgent nature of
these procedures, there is not enough time for tissue typing to be done.
Though tissue matching is proven to increase survival rate of the patient, quickly transferring a
healthy organ to the recipient is more important. Considering that over 75% of organ donations
come from cadavers, time is an important limitation that determines whether an organ remains
viable for implantation. Certain organs, like the heart and the lung, remain viable only 4-6 hours
after blood stops circulating3. Traditional technologies to perform tissue typing include
serological, cellular, and molecular tests. Current tissue typing takes around three hours and
requires the expertise of trained lab technician, meaning the process cannot be started until the
organ has arrived at the laboratory. The (1) inconvenient location of the lab and (2) time needed
for the diagnostics test make tissue typing techniques impractical for most organs, two
limitations ImmunoFluidics overcomes.
Patient-Donor Matching during Organ Transplantation - Issues:
 Currently impractical due to time needed to tissue type organ
 Incorrect typing contributes to $1.5 billion dollars in cost every year
 Limited to diagnostic tests available in central laboratories
 Need expensive laboratory expertise
 Currently only practiced on kidney (longer explanted survival time) and bone marrow
transplant (live patient)
ImmunoFluidics directly competes with other platforms available to perform PCR based
diagnostics tests. Current microfluidic PCR technologies enable different applications of PCR
techniques; however, these platforms still employ inefficient, bulky external heating elements.
ImmunoFluidics relies on novel microfluidic technologies to maintain a sustainable competitive
edge.
Page 2
Competitive Edge
ImmunoFluidics vs. Traditional PCR:
Advantages of Microfluidic Diagnostic Technology:
 Combines faster reaction times with decreased reagent usage.
 Allows portability for point-of-care or point-of-application diagnostics
 Requires minimal expertise due to on-chip automation
 Provides faster reaction times due to high surface area to volume ratio in the micro scale
 Less thermal mass for heating and cooling during biological diagnostic techniques
 Ability to multiplex reactions.
Competition
Competition with ImmunoFluidics would come from other companies attempting to create
microfluidic PCR devices. The following are top companies involved with this technology:
1. RainDance
 Materials: PDMS Chip, pipette tips, drive oil, carrier oil, stabilizer/destabilizer reagents
 Pros: 4,000 regions per sample, highly accurate, no valves or moving parts on chip,
disposable chip, requires picoliter samples
 Cons: Off-chip heating, $225,000 for instrumentation, of-chip preparation, manual
loading of multiple wells
2. Fluidigm
 Materials: Chip created from multiple layers of fused rubber
 Pros: 48 regions for each of 48 samples, computerized loading device for fewer liquid
handling steps, requires nanoliter samples
 Cons: Off-chip heating, 4 hours from start to finish, $79,000 for instrumentation
3. HaloPlex
 Materials: Biotinylated probes
 Pros: Custom designs of probes up to 500 Kb, great specificity, uniformity and high
sensitivity
 Cons: 6 hours for amplification, 96 samples, 480 reactions
Competitors demonstrate complex microfluidic function and manipulation for PCR diagnostics;
however, there are limitations to many of their technologies that ImmunoFluidics will address.
Device Design
Technological developments in the industry have focused on sensitivity and size of PCR
devices. Simply put, PCR is a technique that uses cyclic heating for controlled amplification of
genomic information. Critical aspects of PCR devices are (1) heating control/speed, (2) sample
loading and (3) DNA analysis. Our device presents novel technologies in heating as well as
loading. Further DNA analysis will rely on using previously developed platforms provided by
collaborating companies. Figure 1 represents the general design of our device as well as a
picture of our prototype.
Page 3
Figure 1: general design of our device (left), finished prototype (right)
Design Specifications
Design
Decision
Electrode
Composition
Electrode Shape
Composition of
Base/Electrode
Layer
Specification
Gold – Sputter
Coat
Oven Heating
Coil Concept
Polystyrene –
“Shrinky Dink”
Concept
Loading Layer
Composition
Polyolefin
Flow Layer
Design
Sealing Layer
Simple Loading
of Wells
Polyolefin
Well Number
Chip Size
Rationale
- High conductivity and well characterized resistivity
curves
- Sputter and shrink technology allows for no clean room
-Consistent localized heating achievable
-Best material properties, chemically inert
-Ability to create complex designs
– Allow easy prototyping, fabrication (no clean room), and
scalability
-Low cost, easy to fabricate, adhesive nature makes for
easy manipulation
-Fast and easy fabrication allows for scalable loading
-Low cost, easy to fabricate layer prevents crosscontamination and evaporation during thermocycling
4-5
-Basic design for proof of concept
- Plan to upscale later on
Roughly 3cm by - Increased portability
2.5 cm
Prototype Device
Our prototype device will be loaded by a single syringe with positive pressure. Furthermore, our
device will be powered by an external power source connected directly to the electrode contact
pads. Specific device operation can be seen in Figure 2.
Page 4
Figure 2: Device Functionality and Operation
Device Novelty
ImmunoFluidics directly competes with other platforms available to perform PCR based
diagnostics tests. Current microfluidic PCR technologies enable different applications of PCR
techniques; however, these platforms still employ inefficient, bulky external heating elements.
ImmunoFluidics relies on novel microfluidic technologies to maintain a sustainable competitive
edge. ImmunoFluidics leverages novel patented fabrication techniques, developed in labs at the
University of California at Irvine, to provide unparalleled reaction speeds on a platform that is
biocompatible, cheap, and easily manufactured. This novel manufacturing is patented and
owned by our mentor, Dr. Michelle Khine as well as by the University of California, Irvine. From
our collaboration with other diagnostic companies, we will be able to license and use preexisting
patents from our collaborators at very low costs if any.
Page 5
Sustainable Competitive Technological Highlights
Our device has been optimized and tested for functionality through finite element modeling and
a series of reiterations. Initial testing and has shown promising results that our device can
surpass the competition. Initial electrode heating capabilities have been identified through IR
camera imaging as seen in figure 3. Further analysis and data needs to be gathered to support
the complete validity of our device. We expect full PCR capabilities within the next few weeks.
Page 6
Figure 3: thermal map (top), temperature over time (bottom
References
[1] Bentley, T. Scott and Steven G. Hanson. “2011 U.S. Organ and tissue transplant cost
estimates and discussion.” Millman Research Report. April 2011
[2] S. J. Lee, J. Klein, M. Haagenson, L. A. Baxter-Lowe, D. L. Confer, M. Eapen, M.
Fernandez-Vina, N. Flomenberg, M. Horowitz, C. K. Hurley, H. Noreen, M. Oudshoorn, E.
Petersdorf, M. Setterholm, S. Spellman, D. Weisdorf, T. M. Williams and C. Anasetti, Blood,
2007, 110, 4576-4583.
[3] Abdulla K. Salahudeen, Naeem Haider and Warren May. “Cold ischemia and the reduced
long-term survival of cadaveric renal allografts.” Kidney International (2004) 65, 713–718