[Histonet] RE: freezing artefact holes on edge of cortex in mouse brain sections

[Aine Behan]

Hi all,

 

I was looking for info on troubleshooting what I think is freezing artefact
holes (FA) in our tissue sections. It is quite specific to the edge of the
brain sections predominantly around the most superficial layers of the
cortex and most obvious in the most lateral aspects of the cortex in coronal
sections.

 

Brains were fixed in 4% PFA following cervical dislocation and dissection.
They were fixed in graded sucrose concentrations of 10%, 20% and then 30%
over 3 days followed by rapid freezing in isopentane and dry ice. These
brains are stored at -80 until cryostating coronal sections (10 microns
thick). The deeper brain structures are perfectly intact and free from FA
holes.

 

Has anyone observed this before and do you think this is FA or something
else? I can send on a H&E pic if it proves helpful. It might be a bizarre
question but could there be anything to do with the cryostat that could be
causing this!?!

 

Regards,

 

Áine

 

Áine Behan, PhD

Department of Psychiatry,
Royal College of Surgeons In Ireland,
Smurfit Building,
Beaumont Hospital, 
Dublin 9. 

Ph: +353-1-809-3857/3798 
Fx: +353-1-809-3741