Chirayu,
Usual problems with heads of late mouse embryos is poor dehydrations. My
recommendation is to use isopropanol instead of absolute ethanol.
My protocol that worked very well for many years was:
Fixation with NBF for 4-6h, short rinse with PBS and storage at 70% ethanol
overnight
(Alter
Chirayu:
I think there are several issues in your protocol:
1- the fixation time looks well but when you wash the fixed specimen overnight
with running water you "unfix" the tissue because formalin fixation is totally
reversible. You end with an unfixed tissue. After fixation do not wash the
fix
Chirayu
a happy new year to you.
I am guessing that there could be 3 possible problems.
1. It appears that the xylene is not changed - maybe the xylene should have
more than one change to eliminate all the alcohol.
2. The alcohol you are using as 100% may have some traces of water in it.
3. Th
Hi,
I think that your problem is not an enough infiltration time into Xylene but It
is a not complete
dehydration of the specimen quite thick.
When it happens I come back to new absolute ethanol and then again into Xylene
up to complete
clearification.
I process small pieces so my scheduled t
