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Electrophoretic Tissue Clearing (ETC)
0.5% acrylamide concentration, no polymerization?
Has anyone tried embedding with a 0.5% acrylamide concentration? I just attempted, but its still just pure liquid. Is this normal, or did I mess up the degassing perhaps?
did you add bis acrylamide in your hydrogel monomer solution?
I try the A4P0(4%acrylamide,0%PFA,0%BIS) and A2P0(2%acrylamide,0%PFA,0%BIS)for 3h in 37℃ water bath,and none of them polymerized.
Yes, I had 0.5% bis acrylamide and 0.5% PFA. I put them in a 37c heating cabinet. I increase or reduce all the ingredients proportionately. But apparently, after I played around with it, 0.5% is just not enough for it to make a gel. The brain tissue still polymerizes, weakly, but I find 0.5% to be a bit too low. 2% PFA 2% bis acrylamide works perfectly. Much recommended. 4% takes too long to clear imo.
Hi Lijun, i also tested A4P0 and couldent see any polymerization beside of the tissue. In the paper they mention that this concentration will only polymerize in the tissue due to the PFA inside the Tissue.
So far i got pretty good results with this Embedding Soution.
, when you say that your brain tissue "weakly" polymerizes and that 0.5% is too low, how do you see this? I use 4%PFA & 1-0.5% Acrylamide, with no polymerization outside of the tissue, but with good results in terms of tissue morphology, antibody staining etc.. No indication (as I can see) of unsufficient polymerization of tissue.