Transcript Presentation by Matthew Wheeler

Microfluidic Technology
for Assisted
Reproduction
Matthew B. Wheeler1 and David J. Beebe2
1Department
of Animal Sciences and Beckman
Institute for Advanced Science and Technology,
University of Illinois at Urbana-Champaign;
2Department of Biomedical Engineering
University of Wisconsin-Madison
Acknowledgements
Eric Walters and Sherrie Clark @ UIUC
Henry “Gripp” Zeringue @ UW
Lorraine Leibfried-Rutledge @ Bomed
Kathy Haubert @ Vitae LLC
Funding Sources: CFAR, NIH, UIUC, USDA, UW
Why micro?
Micro Bio Fluidics
• surface-to-volume ratio is 20,000 for 100 µm dia.
• Reynolds number small
• “2 phase” - cells, molecules, embryos
For more see:
E. M.Purcell, Life at Low Reynold’s Number,
American Journal of Physics, 1977
Low Reynolds Number
Flow
inertial
Re VL

viscous
Turbulence
Improved manipulation?
More in vivo-like?
Microfluidics
Applications
• MIT technology review (2001) - “one of ten
technologies that will change the world”
• Markets
–
–
–
–
–
–
Point of care diagnostics
Discovery/screening (not just drug)
DNA manipulation and processing
Analytical instruments
Drug delivery
Sensing
– Assisted Reproduction
– Bioproduction
– Chemical engineering
– Chemistry
A microfluidic chip, fabricated
from silicone elastomer, that
contains 2056 integrated
microvalves in an area of one
square inch. The chip is
analogous to an electronic
comparator and is an example of
microfluidic large-scale
integration. The complex
plumbing in the chip allows 512
chambers to be mixed pairwise,
with individual addressing and
recovery of the results. [Photo: S.
Maerkl]
Microfluidic Large-Scale Integration
Thorsen et al. Science 298: 580-584.
Rationale
Handling Steps in µchannel
IVF
Handling Steps in Conventional IVF
Oocyte
Load oocyte/embryo
1st Rinse
1st Rinse
2nd Rinse
Supp lements
IVM
(Glucose,
3rd Rinse
2nd Rinse
Hormone s)
1st EC Medium
IVM Medium
1st EC Medium
1st Rinse
2nd EC Medium
3rd EC Medium
2nd Rinse
Remove embryo
IVF
3rd Rinse
Insem. Medium
EC
Change medium
and/or add
Sperm, etc.
Sperm
Blastocyst
Blastocyst
Hardware
Inefficient & labor
intensive
Automate procedures and
improve efficiencies
Traditional
µFluidic
Device Design
• Device Design
–
–
–
–
Funnel - loading & unloading
Parking place - holding/placement
Channel - microenvironment
Wells - reservoir
Micro Engineering
• Computer chip (IC) manufacturing
– Lithography, deposition, removal (etching)
– Silicon, metals
• MEMS (MicroElectroMechanical Systems)
– IC methods to make mechanical things
• Micro Fluidics
–
–
–
–
Borrowed from above to make small pipes
Largely glass & polymers
Caliper (Capillary Electrophoresis)
Micro arrays (Nanogen, Affymetrix)
Basic Logistics
1. Loading/unloading
2. Transport (no cumulus)
3. Transport (with cumulus)
QuickTime™ and a
Sorenson Video decompressor
are needed to see this picture.
Chemical Manipulation
Zona Removal Device
Before
After
“Parking place”
7.5 m
200 m
200 m
IVP (mice & pigs)
• General conditions
– Static (“no flow”)
– Straight channels (250µm high x 1000 µm wide)
• Demonstrated
– IVM, IVF, EC
Mouse Culture (Development)
% Blastocyst
100
90
80
70
60
50
40
Microchannel
Control
30
20
10
B6SJL/F1 x ICR - less vigorous
0
24 h
48 h
72 h
96 h
Mice, Pigs and Cows
What’s really going
on?
Micro fluidics
Environmental
control
Reduced
“effective” volume
Micro environment
Micro Fluidics
• Effective volume
– ~50 µl vs. ~ 50 nl
• Environmental control
– Rapid, precise stimulus application
– Gradual, gentle media changes
• Micro environment
– Diffusion governs transport
– Good & bad stuff hangs around
Opportunities for Nanotechnology in
Agriculture and Food Systems Research
1). Food and water supply monitoring:
-presence of residues, trace chemicals,
antibiotics, pathogens, toxins);
- integrated, rapid DNA sequencing to identify
genetic variation and GMO’s;
-integrity of food during transportation and storage
2. Animals health monitoring:
-developmental biology;
-presence of residues, antibiotics, pathogens, toxins;
-bio-sensors
3. Environment monitoring:
-land, water and air pollution;
-remote/distributed sensing
Objectives of a National
Research Program
1). Develop agriculture and food systems related
microfluidic devices that feature integrated
operations, simple reliable components and
low costs.
2). Integrate microfluidic devices flawlessly
into a wireless-ID network.
3). Others ? ? ? ?
How is Agriculture Different?
(or how will USDA’s focus differ from DOD, NSF, NIH or NASA)
1).
2).
3).
4).
The users are producers, processors and consumers.
-applications must be simple, reliable and
highly accurate
The “environment” is dirty.
-samples need some processing (filtration,
purification, etc.)
Agriculture and Food Systems are highly integrated.
-applications need to be networked and results
integrated
Single sensor applications are likely not inadequate.
-fields, herds, flocks, elevators, trains, trucks,
processors, manufacturers etc. are widespread
Outcomes and impacts should address these issues!
Research Budget Estimate
1). Biochemical/Genetic/Residue detection
-$5 million/year
2). Systems for high-throughput drug/cell
screening and biosensor applications
-$5 million/year
3). Automated/integrated networks
-$3 million/year
4). Application testing
-$3 million/year