[Histonet] Leaving sample over night in ETOH 70% during fixation
Itai Moshe
itai.moshe <@t> mail.huji.ac.il
Thu Jul 7 03:22:16 CDT 2011
Dear All,
Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
during the fixation process, will be better for the tissue fixation, and
does not harm the sample ?
Does the fixation process should be done straight forward from the first
step to the last one without any over night stops (except from the PFA,
Bouin's step) ?
My fixation protocol is like this:
1) immediately after killing the mouse i'm putting the sections in a
fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
DDW - pH 7, Or bouin's solution over night at 4C.
2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
3) Xylen x2 - each for 1Hr at RT.
4) Paraffin x3 - each at 60C for 1Hr.
Thank you all very much in advance
Itai M
Doe
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