This week, Christine and I have been trying to figure out if we can measure transport in the Tecan with the transwell inserts remaining within the 24-well plates. Here is a schematic of our idea (with sodium fluorescein, Na-F, as the model fluorescent molecule for transport studies): The left image is a side view, the…
Christine and I have been using the present Corning 24-well plate definition in the Tecan for our experiments so far. Yesterday I placed a pnc-Si chip in the (approximate) middle on the bottom of 1 of the 24-wells. I did a 500nm absorbance scan in a circular pattern (15×15 points) – to get data similar…
Jim and I were wondering if we could “image” the membrane windows with the Tecan. A nice thing about the Tecan is that you can specify the number of measurements per well. For example, in this study, I acquired a 10×10 matrix of fluorescence intensities in a “filled circle” pattern (see below). You can also…
I recently completed a HUVEC adhesion study with commercial PET and pnc-Si transwells. HUVEC were seeded @ 100000 cells/cm^2, allowed to attach for 5 hours (upside down format) and then stained with CMFDA. Images were acquired without autocontrast, but I did autocontrast them in ImageJ to make them brighter. Corning Costar PET transwell: pnc-Si transwell…
Since HUVEC morphology hasn’t looked good in short-term adhesion assays recently, I analyzed HUVEC on the 2nd day after cell seeding. Normally, CMFDA staining is done for 30-45 minutes in serum-free media. I wondered if the serum-free environment during staining was affecting the HUVEC. So, I stained one sample with 5uM CMFDA in serum free…
This was my first experiment in which the Sepcons were inverted for cell adhesion. After allowing the cells to attach for 4.5 hours, I inverted the Sepcons in media for staining with CMFDA. Therefore, the Sepcon is set up so that the pnc-Si side is down and the cells are “upside down”. This makes the…
This is a continuation of the last post on AFM analysis of pore changes during discoloration. I took the AFM images, inverted them in ImageJ so that the pores would be white and then did PoreProcessing in MATLAB. I simply selected a threshold but did no binary operations. The calibration for these AFM images was…
To complement the recent study which investigated pore changes during discoloration with TEM, I did AFM on the same samples. These were w673 samples which were incubated in DMEM (-FBS) in the 37C oven. We didn’t use FBS because I thought the serum proteins would have adsorbed to the samples and then made AFM analysis…
Chris gave me a few wafers to look at with AFM – an oxide sputtered in the recently cleaned chamber and VA Semiconductor wafers cleaned with “Normal Power” or “Full Power” Megasonics. I employed a couple of new “tricks” to help with the imaging this time. All of these scans were taken almost entirely in…
Lately, Anant and I have been doing our discoloration experiments under purposely harsh conditions so we don’t have to wait for multiple days for experiments to finish. We have lots of evidence that the 3D format slows down discoloration (post1, post2, post3). This experiment was to test how long samples will last in 3D format…
