A droplet (3uL) of 0.117 uM actin solution was added on membrane W303(0,2). Let the membrane sit in humidity Chamber for 20 min. Remove excess liquid. Add equal volume of Uranyl Acetate on membrane. Wait 90 sec. Remove excess liquid. Image was taken right after sample preparation.
Xiao
In trying to determine the best conditions for capturing exosomes, one of the important questions that we have is what produces the case where exosomes are caught directly in the pores, not on top, but flush with the surface. This should be the size where the particles are almost exactly the size of the pores,…
Since HUVEC morphology hasn’t looked good in short-term adhesion assays recently, I analyzed HUVEC on the 2nd day after cell seeding. Normally, CMFDA staining is done for 30-45 minutes in serum-free media. I wondered if the serum-free environment during staining was affecting the HUVEC. So, I stained one sample with 5uM CMFDA in serum free…
JP handed me a stack of wafers today, and I went over to the Krauss Lab and with the help of Jack Calcines got some AFM images. I didn’t have much time to play with, so I tried to get the most data I could quickly. I think we’ll have to do a more exhaustive…
Previous Post on Water Permeability Figure We decided at the last meeting to show water permeability for each membrane used rather than plot for a specific characteristic like porosity. I did this but there were a few samples that still had a noticeable deviation between experimental and calculated permeabilities. I asked Dave to reprocess the…
Jess showed me how to use the diffusion chamber to separate proteins on the membranes. I ran a gel to see if any proteins appeared in the filtrate. The gel is posted below: The lanes are (from left): protein standard, control, retentate 1, filtrate 1, retentate 2, filtrate 2. It appears that protein diffused through…
Are you wondering why Dean has yet to go beyond 30% BSA in his clearance studies? Here is why … APP-04_Viscosity-of-BSA-in-PBS.pdf
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There appears to be molecular structure in these images! In my experience people need to label with larger actin binding proteins to see this type of structure. I know Karen is doing some comparisons on carbon grids.
Could be good.
Here is a useful web page on electron microscopy that uses actin filaments as a reference molecule. The good news is that I think our images show hints of the ‘rope-like’ actin structure you can see in Figure 1.8.
http://www.mih.unibas.ch/Booklet/Lecture/Chapter1/Chapter1.html