I repeated a previous experiment but this time I used samples (no RTP) from W685 in oven @ 37C in DMEM(-FBS). Four type of samples (blank chip with no slits, chip with both intact membranes, chip with one intact membrane and chip with both broken membranes) were used at different time points. Again the expected…
We have known, W683: no visible pores; W684: very few pores; W685: usual porosity. After Vlassak’s view on discoloration, I took samples from above wafers to check if any difference in discoloration can be seen as they have different porosities. We could expect the more porous material to degrade faster than the less porous and…
Last week when I reported no membrane breakage after a week of HUVEC culture, I showed a phase micrograph on day 7. This week I tracked the HUVECs a bit more closely with the microscope. This is 10X, HUVEC P5, initial seeding density of 32,500 cells/insert. I tried to capture approximately the same area on…
Previous Post The previous w670 pressurized flow gels showed one trial with a lower cutoff than w673 and one trial with nothing passing through. This experiment was run for 40 min and in each case I saw only 0-3 uL fluid passing. I used the last couple of samples from this wafer to attempt a…
I recently completed some experiments with the Endohm system, which measures transendothelial electrical resistance (TEER). I seeded either 16.25k or 32.5k HUVECs (P5) on commercial polycarbonate (PC) or polyester (PE) transwells and on pnc-Si in the transwell format. I then tracked the TEER for 7 days. This data is normalized to the day 0 TEER…
Since we’ve been noticing slower discoloration in the Sepcon format, I looked at the 2 components of Sepcons – the plastic and the O-ring. Specifically, I cut pieces from w622 and incubated them in FBS-DMEM/F12 either alone, with the Sepcon plastic or with an O-ring. This was in the oven, and the chips were membrane…
In an earlier post we saw that intact membrane sample discolored slower than the broken one. To establish this, a study using four types of samples from #702 (blank chip with no slits, chip with both intact membranes, chip with one intact membrane and chip with both broken membranes) was carried out. The samples (no…
I carried out the bead assay to monitor membrane integrity in Sepcon format. The sample (no RTP) was taken from #703 and put in DMEM(-FBS) @ RT with membrane side facing up. 15um polystyrene beads were allowed to settle on the membrane and images were taken. As can be seen, the membrane is still intact…
XTT is a colorimetric assay for measuring cell viability. Supposedly, it gets taken up by cells and is reduced by mitochondrial activity intracellularly into water-soluble orange formazan crystals. The readout is the absorbance of these orange crystals. I used XTT to get standard curves of HUVECs in 24-well plates first. Here’s the data from 2…
Based on Barrett’s array set up for discoloration, I set up an array experiment to check the discoloration effects between the new and old system ( 2e-7,AJA vs the older 7e-7 CVC ). The wafers from each had a bit of the wafer with different treatments (900C RTP,1000C RTP and Amorphous). To incorporate the maximum…