This post presents the data from all of my TEER experiments of BBB co-cultures, with bEnd3 endothelial cells on the bottom and NG101815 glia on the top of PET and pnc-Si transwells. For all experiments, I used 50000 cells/cm2 as the seeding density. I always had 3 samples – bEnd3 alone, NG10815 alone and the…
A few weeks ago, I posted some high resolution (100X) images of BBB co-cultures stained with 3 different colors. I also included the z-height of the different focal planes of cells on either side of PET transwells. In this post, I present a repeat of this experiment, along with the same analysis performed with cells…
To follow up my last post on air-blood barrier TEER results, I wanted to show some Live/Dead images of these samples. These are stained after the 14 day experiment. In the following image, you can see normal bEnd morphology on PET and pnc-Si. Only the cells over the pnc-Si slit are visible because these cells…
I finished 3 independent experiments of air-blood barrier co-cultures – these consist of bEnd3 endothelial cells on 1 side and RLE-6TN lung epithelial cells on the opposite side of the membranes. We didn’t expect these cell types to potentiate TEER in anyway (like we expected glial cells to tighten endothelial barrier function). However, our collaborators…
As a continuation of the substrate bias series, I deposited a 15 nm film with 05 W substrate bias and annealed at 1000 C. TEM shows that this condition generates a 13.4% porosity and a max pore size of ~40 nm! Here is a comparison of all the bias conditions tested thus far: I believe…
Last week, I reported on the the morphology of pnc-Si deposited with different substrate biases. We hypothesized that the reason for this was due to the different initial film stress of the as-deposited a-Si. Unfortunately, the pressure-displacement method does not allow us to measure amorphous films since it requires the membrane to be nearly flat…
Hi everyone! I’m sifting through the mass of data I collected during my last month and am compiling posts to update everyone. This post shows the latest with the microfluidic chamber that Henry and I designed. In my first post regarding this device, I showed microparticle and blood flow over cultured endothelial cells (bEnd3). In…
It seems that pores are only formed in the “sandwich structure” on certain process. So in order to verify this conclusion, I annealed two layers structure (20nm sio2+15nm si) to test the agglomeration theory. The other parameters in depositing and annealing are same to typical sandwich structure process (1000°C anneal, ramp rate 100c/s, w/o bias,…
UPDATE: A look at long and short crystal dimensions (9/1/10): DLS vs. Crystal Sizes – Newest version 8/30/10 Here is a comparison of sieving curves using DLS or estimated crystal dimensions of proteins. DLS: 24 hours sieving curve. 48 hours sieving curve. Estimated longest crystal dimension: 24 hours sieving curve. 48 hours sieving curve.
We have always speculated that the substrate bias during the a-Si deposition had a strong effect on pore morphology of pnc-Si. Here, I looked at three 15 nm silicon films deposited with 00W, 25W, and 50W substrate bias. Recall, that the substrate bias applies rf energy to the substrate surface so that impinging Si atoms…