[Histonet] Flash-freezing samples in OCT?

[Greg Dobbin]

Hi Mariano,
I think histologically the tissues will look fine. However, snap freezing by submersion in N2 quite frequently causes the tissue blocks to crack which may present some mild sectioning issues. 
Greg


Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE    C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385


>>> "Mariano S. Viapiano" <msviapiano <@t> yahoo.com> 7/6/2007 7:40:10 PM >>>
Hi everyone, 
We just received some frozen brain tumor samples to
process for IHC but are a bit confused about how they
were prepared. We fix our samples in 4% PFA, change
them to 30% sucrose until they sink, embed in OCT and
freeze them at -80C until cryostat sectioning. On the
other hand, the samples we received were fresh
specimens (just out of the OR) dipped in OCT and
snap-frozen in liquid nitrogen. What would be the best
thing to do? Section them as they came, or thaw them,
remove the OCT and do our fixation procedure before
sectioning? I am a bit concerned about the OCT having
acted as a “retardant” for the snap-freezing of those
samples when they were submerged in nitrogen (or am I
just imagining things?)
Thank you!
Mariano



Mariano S. Viapiano, PhD
Assistant Professor
Center for Molecular Neurobiology and
Department of Neurological Surgery
The Ohio State University
226B Rightmire Hall
1060 Carmack Rd., Columbus OH 43210
Tel (614) 292-4362
Fax (614) 292-5379


       
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