In my last post on HUVEC intercellular junctions, I just showed cadherin staining. I repeated this protocol but also tried immunofluorescence for ZO-1, the tight junction protein. These are HUVEC P7, 6 days post-seeding @ 50,000 cells/cm2. I looked at…
Category: NRG
BSA-Au Separations
The BT method mentioned in this post seemed to give stable particles, but the BSA-Au did not work in separations. After about 24 hours, the particles retained pink color but seemed to settle in tubes. This may be due to…
Posted in NRG
HUVEC Cadherin Immunofluorescence (1)
One of the markers of endothelial monolayer maturation/barrier function is the expression of adherens junction and tight junction proteins. The adherens junction is composed of transmembrane proteins called cadherins (among other proteins) that link cadherins of neighboring cells to the…
Posted in NRG
HUVEC Mono(?)layer Na-F Permeability
I had some old HUVEC (P8) on pnc-Si transwells and PET Costar transwells that I didn’t know what to do with. So I tried a permeability study to see if there were any unexpected roadblocks in the protocol. I seeded…
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Spinning transwells
In my previous transport post here, I showed O-ring placement data which was motivated by my thinking about the unstirred water layer (UWL). I think it’s helpful to think about the total resistance to transport through transwell membranes as a…
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Fluorescein transport: O-ring position and wafer variability
After diffusing through a polymeric or pnc-Si membrane in the transwell format, diffusing molecules meet an “unstirrred water layer = UWL” on the bottom of the membrane. The UWL acts as another resistance to transport into the receiver compartment. According…
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Dilution Problem
This was mentioned in the meeting today, and I’ll try to describe what’s going on here. We start out with a system with a smaller volume on the top of the membrane: If we start out with 10 mg/mL in…
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Stock Pile Experiment
This experiment is to determine if the time between the SC1 clean and when the wafers get deposited matter. Currently there is suspicion that an SC1 from an RCA clean should at RIT should be done right before deposition, .…
DNA Filtration at New NaCl Concentrations and New Salt Types (LiCl and CsCl)
I ran new experiments over the weekend in which I tested NaCl at 10, 25, 60, and 155mM. I also ran filtrations in the presence of 10mM LiCl and 10mM CsCl. These experiments were performed with chips from wafer #…
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Particle Separations with Cellulose Membranes
I tried particle separations with the 30kD cellulose membranes from Millipore using the same setup as previous experiments. Basically the results are that neither 5 nor 20nm particles pass through the membranes whether in water or 10mM KCl. Absorbance Data:…
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