Combining Chemical and ET Clearing
KatherineHolzem
Posts: 21
So, I stumbled on this by accident, during my staining, but found something that might work well. Basically, I was attempting a TSA staining, which requires a peroxidase blocking step (basically H2O2 incubation). After this step, the tissue went from having a bit of color and opacity to being completely clear and colorless... I could hardly find the sample.
I know that H2O2 is basically the Dent's fixative in chemical clearing procedures, so I was wondering whether many of these procedures could be improved by combining chemical and ET clearing. I didn't clearly notice any ill effects of the H2O2, but then my staining didn't really work either (which could have just been the antibody). Definitely had autofluorescence though.
I know that H2O2 is basically the Dent's fixative in chemical clearing procedures, so I was wondering whether many of these procedures could be improved by combining chemical and ET clearing. I didn't clearly notice any ill effects of the H2O2, but then my staining didn't really work either (which could have just been the antibody). Definitely had autofluorescence though.
Comments
Embryos for Ultramicroscopy
Klaus Becker, Nina Jährling, Saiedeh Saghafi, and Hans-Ulrich Dodt
In this paper they also use the peroxide/Dent's fixative for clearing (but of mouse embryos). However I doubt this protocol would be useful since it involves dehydration and that would severely reduce the fluorescence.