Hello!
I am looking for the working protocol, which I could use for clearing the brain section, 105 µm thick (hemisphere).
Without this I am not able to reach deeper into the tissue than 20 µm,with confocal microscope with Airyscan, what is problem here. Important thing is, that our section has GFP produced by animal, so crucial thing is to have GFP signal in cells saved.
That is difficult. You can try this protocol after mounting the sections onto a slide : 5-10 dipps in toluol or xylene, coverslipp them with DPX.
If the sections try to curl put the slide into a petri dish, cover it with a filterpaper soaked with toluol or xylene and put a small petri dish on that sandwich. After 5 min remove everything and coverslipp the flat sections with DPX.
The disadvantage is that the section shrunks in the dorso/ ventral direction because of the solvents and xylene solublee DPX.
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