Combining Chemical and ET Clearing

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.


  • irinairina Posts: 6
    Immunostaining, Dehydration, and Clearing of Mouse
    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.
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