This is some preliminary data that I put into my F32 postdoc fellowship application. It is a comparison of rhodamine transport through pnc-Si and PET transwells. These studies were done with PBS as the buffer. Sample volume = .1mL, donor volume = .3mL, receiver volume = 1mL, initial [rhodamine] = .5mM. I set the experiments…
There were some ideas being thrown around a couple weeks ago about discoloration results. I tested a couple of them out last week. These experiments were with w615 pieces, in the 37C oven in serum-free DMEM. I tested if vacuum grease or nailpolish would cause discoloration, I repeated the membrane up vs. down experiments that…
Recently, I’ve been frustrated with polycarbonate transwells since they are opaque and I couldn’t see cells on them. So, I bought new transwells from BD (Falcon, PET membrane, 0.4um pores) and decided to test these out. I also wanted to see if coating the transwells with gelatin would promote cell adhesion. These are P6 HUVECs…
We’ve been wondering about the effects of membrane stresses on discoloration rates. This was spurred by Vlassak’s visit and his ideas about stress and membrane stability. In this experiment, I used w501, no RTP, in the oven in RPMI+10%FBS. All of these samples were in the Sepcon format: chip sealed with a fat O-ring, chip…
Last week, I repeated Anant’s experiment which aimed to eliminate edge effects by painting over chips with nailpolish. I painted the entire well side of the chip but tried to keep the wells clear of polish (“painted back”) and I covered just the edge of a chip but tried to keep polish off the well…
I got a couple of wafers from Chris last week – 50nm oxide deposited at room temperature or 450 degrees. For each wafer, I saved the zsensor (top), phase (middle) and zsensor (bottom) scans. I also calculated the roughness (the table looking thing on the bottom of each image. I had some problems with imaging…
I wanted to do some more fluroescent imaging in transwells to look into background fluroescence. This was mainly motivated by my inital study. These are HUVECs stained with 5uM CMFDA and 10uM Hoechst 33342, fixed in formaldehyde and imaged on the Zeiss at 10X. This was 6 hours after seeding. Before acquiring the images, I…
I did an experiment earlier this week in the Sepcon and flat format. I intentionally broke membrane windows so that I could compare chips with no intact membranes vs. chips with 1 intact membrane (and also the effect of autoclaving). Unfortunately, all of the flat chips discolored overnight, so I couldn’t see any differences between…
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…
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…
