Carrie and I have been trying to detect HMM (APOBEC3G) diffusion through Sepcons without much luck.  We determined that the dilution factor (~1:30) for diffusion in our 384-well plate format was too high to detect HMM in the basolateral well.  To try to get around this problem, I decreased the volume (and thus dilution factor)…

A couple weeks ago, I presented a potential figure for my paper that didn’t make sense.  So, I did a couple more controls and put together this new figure.  This is the “diffusion through ‘non-porous’ SC348/cut-off of nonporous membranes” figure.  I redid it to include SC348 (the nonporous sample) and ‘typical’ pnc-Si as % of…

In the soon to be submitted paper on diffusion models, we list two contributions to membrane resistance.  The first is due to the steric and frictional hindrance a molecule encounters as it enters and diffuses through a pore (Renkin, Deen).  The second is due to the time that it takes for the molecule to diffuse…

This post is related to the HIV drug screening project that Carrie has been working on.  To refresh everyone’s memory, the aim of this project is to use pnc-Si as a filter to separate high molecular mass (HMM) protein-RNA complexes from smaller protein monomers/multimers.  Small molecules that break down the HMM form of this protein…

I first reported the crystallization of free-standing amorphous silicon membranes in this post in January. The last experiment was done inside the susceptor (top and bottom piece). This week, I annealed a set of amorphous 15 nm and 30 nm with the susceptor top off. This allowed for the direct heating of the films. One…

As a follow up to my 15 nm post-Si deposition Ar sputter experiment, I fabricated a set of six wafers with 30 nm Si films this week: three with the Ar sputter process and three without. I annealed these at 1000, 1050 and 1100 C with a ramp rate of 100 C/s for 60 s….

The mechanism of organic dye photobleaching is not well understood, but there are a few methods out there for reducing the rate of photobleaching.  Some literature suggests that photobleaching is due to excess oxygen in solution and the interaction between the dye and oxygen radicals.  Actin research swears by the following “anti-bleaching buffer” to limit…

This is simply the latest repeat of a blood-brain barrier co-culture on PET and pnc-Si transwells.  These are P17 bEnd and P24 NG10815, each seeded at 50000 cells/cm2.  As usual, I seeded bEnd on the pnc-Si side, upside down, and NG10815 on the well side, facing up.  I took TEER measurements every other day for…

In most of the images I’ve posted of bEnd3 on pnc-Si, it seemed like there were more cells crowded onto the free-standing pnc-Si compared to supported pnc-Si.  This post shows that this is indeed true. To do cell counts, I stained cells with Hoechst 33342 (1uM in media) for 15 minutes @ room temperature and…

All of my recent posts on VEGF, VEGFR2 and NF-H imunofluorescence have included data from cells grown on coverslips only.  This was my first attempt to try to do immunofluorescence staining on intact pnc-Si transwells and then pop the chips out of the transwell plastic in order to image at 100X.  Since VEGF is involved…