Endothelial cells on small square membranes

I designed new masks with different sized squares and slits in order to study how the aspect ratio of free-standing membranes would affect endothelial cells.  In this 1st experiment, I seeded bEnd at 50000 cells/cm2 and let them grow for 4 days.  I then stained the cells with Live/Dead and overlaid the phase channel in blue to show where the membranes were located.  I’m not including all of the different sized squares in this post – just the 2 smallest designs.  These results are interesting because it (almost) shows vacuole expression at a single cell level.

In the image below, about 2 cells are growing across each membrane (these are the smallest square membranes I had made).  It looks like those cells on the membranes have vacuoles.  Also, some cells that are partly over the membranes expressed vacuoles on supported pnc-Si.
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These are slightly larger membranes (~5-6 cells per membrane).  On these membranes, vacuole expression seems to be limited to free-standing membranes only.

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