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…
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…
My last cell post involved pictures of HUVECs after 1 day of growth on commercial transwells and pnc-Si Sepcons. Over the past week, I’ve been growing HUVEC on commercial PC transwells and pnc-Si in the Sepcon format in order to track the TEER. To get more data out of these cells, I decided to do…
I repeated the autoclave experiment from earlier this week, but this time I also used the Sepcon format. I used wafer 662, which was contaminated with large spots. This was done in the oven in RPMI-1640 with FBS (no RTP). I think my 0 hour pictures were accidentally erased from the camera, but I think…
My previous discoloration post showed that chips in the Sepcon format discolored much slower than flat chips. A flat chip in the same well as a Sepcon chip seemed to accelerate discoloration. One thing about the Sepcon format that I hadn’t thought of earlier was that pnc-Si in Sepcons is autoclaved for sterility. So I…
I’ve been doing some preliminary studies with HUVEC in commercial transwells and on pnc-Si in the Sepcon/transwell hybrid format. The thinness of pnc-Si will enable better microscopic examination of cells than other membrane systems (if pnc-Si is flat). I thought it would be interesting to compare commercial transwells to the pnc-Si “hybrid transwell” format in 24-well plates via…
I ordered some new AC160 tips last week from Asylum and got on the AFM Thursday to test them out. I think they solved some of the problems I was running into over the last few weeks. Mainly, I was finally able to image entirely in repulsive mode (which was impossible with the older tips). All of…
I performed a discoloration experiment with w639 in EGM + 10% FBS. I put 2 blank chips in the Sepcons membrane side down. I then placed one Sepcon in a well with EGM alone. The other Sepcon was placed in a well with EGM but also with another chip (labelled as Sepcon with chip, chip…
This is a repeat of the previous array experiment, but with a higher NaHCO3 concentration (1.85 mg/mL). Also, since these experiments require a lot of samples, I decided to cut chips off of wafer skeletons. This one was w622. As before, these were not RTP’ed and they were placed in the oven at 37C. I…
Based on last week’s lab meeting, I made a “solution array” to test discoloration. Specifically, I made 3 solutions of NaHCO3 (0, .23125, .4625, .925 mg/mL) and 3 solutions of NaCl (0.1, 0.5, 1 M). I placed these non-RTP’ed w648 chips in the oven at 37C and took pictures at 4, 18, 72 and 120…