[Histonet] Embryons fixation and processing
ana.merino-trigo <@t> wanadoo.fr
Mon Jan 23 15:20:03 CST 2006
I just began a new job in a different field. Before I was working with mouse xenograft tumors and now I moved to develpment biology field. I need to process zebrafish, chick, axolt and mouse embryos and I was wondering if anyone could advice in terms of fixation and processing times. Mostly of the papers that I got they fix in 4%PFA (I'm use to fix in 10% formalin, and I was wondering if is common with embryos to fix with 4%PFA). In some papers they fix 2 h at RT or 4ºC o/n, when using 4%PFA. Any advice in this sense?
In terms of processing, a guy that did some immunos with embryos here deshidrates samples in MeOH (25%, 50%, 75% and 100%, 5 min each), but I'm use to EtOH deshydratation (1x1h 70%; 2x1h 95%; 4x1 h 100%). Could someone advice in deshydratation times and advantages of using MeOH?
Thanks a lot,
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