Another one of the questions I’ve been trying to answer with regard to endothelial cell vacuoles is whether membrane mechanics or permeability drive vacuole expression.  I’ve thought of several experimental designs to test this that haven’t worked (i.e., using very…

Last week, I annealed a few amorphous 15 nm TEM grids in the RTP.  We have always speculated that the stress from the bulk silicon was a necessary condition to induce pore formation and I wanted to confirm this.  I…

One of the ongoing efforts with endothelial cell culture on pnc-Si is to confirm that the holes in these cells are actually vacuoles.  In my last post, I tried to do this with FITC-Dextran and TMRIA with indefinite results.  I…

Tom recently designed a wafer with 1cm2 chips containing a 2×2 array of 1mm2 square membrane windows that fit on 384-well plates.  I wanted one of these chips to get a preliminary data image for my R21 application.  Instead, I…

In this post I will describe the use of the 3-D computational diffusion model as applied to separations with thick membranes. Experimental:  Track etched membranes were trimmed to fit into round plastic sepcons and were sealed in the traditional way…

My previous 2 posts on co-cultures presented images but I also tracked the TEER in these experiments.  This post shows that data. The top 2 graphs are from the 1st co-culture experiment – the bottom graph is from the second…

In my last post (the first co-culture), I showed that bEnd3 cells exhibited a different morphology when they were co-cultured.  I also decided that I would reverse the orientation of the co-culture in order to better visualize the endothelial barrier. …

This post presents cell images from the first successful co-culture experiment.  I seeded P8 bEnd3 cells alone, P13 NG108-15 cells alone and both cells on PET and SC500 pnc-Si transwells.  For this experiment, I seeded 50,000 bEnd3/cm2 on the apical…

One of the themes in my recent posts has been the presence of ‘vacuoles’ over free-standing pnc-Si and PET membranes. We’ve assumed these are vacuoles because endothelial cells form vacuoles, they don’t stain with live cell stains (and so have…

I’ve been wondering if there were differences in actin/cytoskeletal structure of cells on supported vs. free-standing pnc-Si.  A couple weeks ago, I had a couple of extra pnc-Si transwells so I stained for actin. I followed the actin staining (rhodamine-phalloidin)…