problems with cryostat sectioning of thyroid

From:Derek Marsee

I am working on a project involving rat thyroids which I must section on a 
cryostat.  The tissues section very well, and the overall histology of my 
sections is pretty good.  Unfortunately, the thyroid colloid does not stain 
- it appears that the colloid is being torn or is absent.  I have seen 
other papers where people use cryostats to section rat thyroids, and they 
see nice follicles with intact colloid.  Any suggestions for how to improve 
my results would be appreciated.  Here is my current protocol:

1)      Remove thyroids and rinse in saline
2)      Embed in OCT and freeze in liquid nitrogen
3)      Section at -20 C in cryostat at 10 microns
4)      Store the slides at -80 C until I'm ready to stain (several days)
5)      Thaw 5 min at 4 C
6)      Fix in 4% paraformaldehyde, 10 min at 4 C
7)      Routine hematoxylin and eosin staining


Derek Marsee
The Ohio State University
Department of Physiology and Cell Biology
105 Hamilton Hall
1645 Neil Avenue
Columbus, OH 43210-1218 USA

(614) 292-9243 (lab)
(614) 292-4678 (office)
(614) 292-4888 (fax)

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