In this experiment, I soaked membranes (from either wafer 1064 [150W deposition power] or wafer 1065 [100W deposition power]) in different fluids that are standard use in cell culture applications, or which provide a similar situation as could be found in such (cause of pH change, salts, etc.). Each membrane is made into a single-well…
In evaluating the usefulness of our membranes for cell culture, it is important to understand if and how cell growth is affected by the surface. The desire is that they be consistent with standard culture substrates (i.e. Tissue Culture Plastic (TC)). I performed two growth trials using HUVECs (Human Umbilical Vein Endothelial Cells), one focusing…
In a previous post I had shown the relative fluorescence of various substrates as normalized to glass. 1um beads (either green or red) were placed onto each surface and imaged with constant EM Gain, exposure time, and intensity. Bar graphs were constructed to show the relative fluorescence (red and green were not normalized to each other)….
In this previous post (also some in this post), I had displayed the difference between the 120nm SiO2 microporous membranes and four different iterations of Track-Etched insert membranes in reference to nuclear (DAPI, blue) and actin (Phalloidin, green) stain. I recently performed the same staining protocol on other substrates: High Pore Density Microporous 300nm SiO2, Low…
While performing an experiment to study HUVEC growth rates on various substrates, I observed an interesting phenomena on the oxide membranes. I was using 120nm, nonporous, 600C annealed SiO2 membranes in this trial. The cells were observed at least once per day over a course of 173 hours from seeding. The growth rates and quantitative…
Method: Oxide membranes in some conditions have been shown to wrinkle over time. To study this, three different varieties of SiO2 membranes were observed (1000C nonporous 120nm SiO2, 150W deposited microporous low-density 300nm SiO2, and 100W deposited microporous low-density300nm SiO2). Beads were placed on the top of each membrane and given ~24 hours to settle…
After seeing in previous posts how optically transparent the SiO2 membranes are, I went about seeing if I could quantify this in comparison to the SiN membranes and also CoStar PET track-etched membranes. In order to do so, I ‘seeded’ red 1µm beads (in 1X PBS) onto each type of membrane (the SiO2 and SiN…
I’ve run a few comparative studies with the CytoVu devices that have SiO2 membranes in them. From a previous NRG discussion, here is the comparison of a single location with only the fluorescent images, and that same location but with phase included: In the phase image, the pores can be seen through the cells, especially evident in…
In this trial, bEnd.3 cells were seeded onto three single-well CytoVu devices with SiO2 membranes coated with a 1% Geltrex coating. After ~27 hours, the cells were fixed and permeabilized (the fixing may be only partial though, as the solution used was more than 5 months old). After this, DAPI and FITC-conjugated phalloidin were used…
After experimenting with some different coatings and membrane types in single-well CytoVu devices, here are some representative images of the results: SiN (Nitride) membrane with a Geltrex non-gel coating @ 44hr: 10x: 20x: 40x: SiO2 (Oxide) membrane with a Geltrex non-gel coating @ 42hr: 10x: 20x: 40x: Also, some Live/Dead Assay images were taken on…