[Histonet] Sucrose cryoprection

[Pablo Sánchez Quinteiro]

Thank for your help Gayle,

I am sorry my mail was not detailed enough.

Yes, I am doing frozen sections with a freezing microtome. The animals are
perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same
fixative. Then they are transfered to Phosphate buffer.

I have problems when cutting the tissue. The sections (60 microns thick)
are apparently nice, but just after putting them into the buffer they do
not stay flat and they fall to pieces after gentle touching with the
paintbrush.

I have seen that after sinking in sucrose the samples show a gelatin-like
appearance. They seem 
swollen and they are transparent. Could be this related to a poor fixation?

For immunohistochemistry it is not recommended posfixation times greater
tan 24 hours, but maybe postnatal or fetal material need longer
postfixation time.

Thanks for your interest.

Pablo

>We need more details on exactly what you are doing??????  and what the 
>problems are?  Are you using a cryostat?
>
>At 11:14 AM 9/29/2004, you wrote:
>>Dear listers,
>>
>>I am cutting at the sliding microtome fetal and early postantal mice
>>brains. Could somebody tell me if is it normal that after sucrose
>>cryoprection the pieces of tissue show a gelatin-like appearance? I have
>>lot of problems cutting this tissue.
>>
>>Thanks in advance
>>
>>Pablo
>>
>>
>>
>>
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>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>
>