Study Proposal:In Vitro Cell Adhesion: Whole Blood, Single channel 5.4 mm chips
The purpose of this study will be to compare the cellular adhesion on PEG vs Bare nano porous silicon nitride membranes using the same fluidic devices as the in vitro clearance study.
The devices will need to be disassembled for imaging.
Materials:
- 6 chips, 3 PEGylated, 3 Bare.
- gaskets for 6 devices
- Tygon tubing
- 2 mini peristaltic pumps
- PBS
- DI Water
- Heparinized Whole blood (ovine)
Procedure:
Prime device with de-gassed PBS, 1 hour recirculating
Pump blood for 4 hours, recirculating at the same velocity as the animal model (32 µL/min).
Flush device with PBS, just long enough to clear the blood, 10 min.
Disassemble the devices.
Fix chips with 4% Paraformaldehyde for 10 min at room temperature
Rinse in DI water for one minute.
Image with both light microscopy and SEM.
Measure area of cellular adhesion for comparison between chip surfaces.
Calculations:
0.5 mL/min, 20 channels. 0.025 mL/min per channel. (25 µL/min)
Cross sectional area HD, (938+500)*310/2 = 222,890 µm2
Cross sectional area single, (1138+700)*310/2 = 284,890 µm2
Cross section ratio, 1.27816
Velocity through single slot chip to achieve the same velocity, 1.27816*25 = 32.0 µL/min
So to match the speed of the HD devices, but not the residency time, pump rate should be 32 µL/min.