[Histonet] Mice heads in Paraffin
mote uncle
moteuncle <@t> gmail.com
Thu Feb 16 08:27:54 CST 2012
Hello
This is my first post to this site. I have encountered a problem with
embedding of mice heads in Paraffin (Gurr). Initially with Embryonic day
13-14, the protocol was working properly and the sections were also nice
but with Embryonic day 17-19 I could see even after several hours of
ethanol wash and xylene treatment the mice heads were soft but still I
embedded the samples in paraffin. When I removed the paraffin blocks from
the mold I could see the mice portion to be white, some flecks of wax
coming out of surface and the surface of wax had some cracks. I could not
rule out what was causing the problem.
Protocol I used for E 17-19
After Fixing in EDTA (2 weeks) and putting the mice heads in running water
for 48 hours,
50% Ethanol 3 hours
70% Ethanol 3 hours
95% Ethanol 3 hours
100% Ethanol 3 hours
100% Ethanol 3 hours
Xylene 2-4 hrs (till clearing of samples was observed)
Paraffin 30 mins
Paraffin 30 mins
Paraffin 30 mins
Paraffin 30 mins
Could anybody share me your protocol for Embryonic day 17-19 and Post natal
days.
Regards
Mote
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