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Optimization of T-Cell Trapping in a Microfluidic Device
Group #19
Jeff ChamberlainMatt Houston
Eric Kim
MEMS- MicroElectroMechanical Systems
• Batch Fabrication Processes
• Cell Traps– High-throughput
experimentation– Complex biochemical
analysis– Single cell analysis– Reagent
conservation– Quick environmental
changes
Cell Trapping Basics
Reagent #1
Reagent #2
Cells / Media
Trap arrays:
Our Project
• Maximize trap efficiency by improving upon current trap designs.– Trap Efficiency : maximized number of
traps with 1 cell/trap
Cell Viability ResultsFlow Rate = 100 nl/min
Flow Rate = 250 nl/min
SolidWorks® Rendering of a Single Well
Picture of Well Array
• Square or rectangular shaped well of any depth
• Thousands of mirrored wells in one etching
• Front Surface Mirrors with high reflectivity
• Nearly orthogonal views of specimen
200 um200 um
Background & Motivation
Three Dimensional Image Information May Be Important for Biological Studies
• Chemotaxis• Developmental Biology• Cellular Division• Pinocytic Loading • Volumetric Measurements
Our System is Constructed From Silicon Wafer <100>
Methods: Fabrication
Silicon Wafer Silicon Wafer <100><100>Grow SiOGrow SiO22
Spin Coat Mask Spin Coat Mask LayerLayerPattern with Pattern with PhotolithographyPhotolithography Etch with HFEtch with HF
Remove Remove PhotoresistPhotoresist
Etch with KOHEtch with KOH
Coat with Platinum Coat with Platinum or Aluminumor Aluminum
Cutaway ViewCutaway View
SolidWorks® Rendering of a Single Well
Results: Dictyostilium Extrusion
AA
BB
CC
AA
BB CC
Primary ImagePrimary Image
ReflectionReflection
Primary ImagePrimary Image
ReflectionReflection
Reflection of Reflection of the Reflectionthe Reflection
ReflectionReflection
ObjectiveObjective
Coupling Microfluidics With the Pyramidal Wells
Si Wafer
Cross Section of One Well
PDMS
Flow
Flow
PDMSGlass
Pyramidal Well Dimensions
10 20 30 40 50
25
50
75
100
125
150
depth_min = 3.22728 cell_radius
outside_dim = 3.48504 cell_radius
Assuming:
0.5 base = 1.2 cell_radius
Micromirror Well Dimensions
Cell Type Cell Dimensions (um) Minimum Required Depth (um)
Outside Dimensions for Min Depth (um)
T-cells 5 diameter 8.0682 17.4252
Jurkats 10 diameter 16.1364 34.8504
Dendritic Cells 10-20 diameter 16.1364 - 32.2728 34.8504 - 69.7008
Dicti 10-20 diameter 16.1364 - 32.2728 34.8504 - 69.7008
Myocyte 15 by 100 24.6737 54.9399 by 37.9399
10 20 30 40 50
25
50
75
100
125
150
Future Directions
• Single Cell Design– Desire single cell per well for optimal imaging– Possibly controllably coupled with another
As opposed to…
Future Directions
?
• Single Cell Design– Desire single cell per well for optimal imaging– Possibly controllably coupled with another
Future Directions
• Flow Modeling– Design a more efficient trapping system– Use flow data to design trap design that will keep cell in well
?
Future Directions
• PDMS-mirror bonding– Questions to whether the PDMS will bond to the mirror array
– Use SU-80 and glass cover slip with a PDMS external flow system?