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…
Here are some high resolution TEM images from a SiMPore-UIUC collaborator. Chris says: The high resolution stuff looks pretty cool, especially the zoom-in on the 8nm pore. I the most obvious crystal planes are <111> with 0.314 nm spacing. If you look carefully, some 0.192 nm <110> planes and 0.136 nm <100> planes can also…
We have seen before that in petri dishes, flipped samples (well side up) discolor faster as compared to the ‘membrane side up’ samples. This could be due to local environment trapped below the chip. Hence, I used Sepcon format so that the sample is raised above the floor to avoid any trapped solution below it….
I repeated the experiment with samples from #703. Two samples were in a dish with one flipped over. Again, the ‘well side up’ sample (flipped) discolored earlier as seen in the 24 hour time point in the figure below.
The movable cutoff gels were more or less successful, but were too dirty for publication (see knowledge page). I’ve been trying to rerun them so that they look nicer, but the darker stain that I used has brought out a few new observations. The first gel following this paragraph is the new gel. The second…