IUBio

Floating cryosections

C. C.
Wed Sep 29 12:09:07 EST 1993


 Netters:

  I'm trying to do receptor autoradiography of brain sections (12 micron,
from a cryostat) but fragments of the sections come off during
preincubation.  The brains are frozen in isopentane at -40C, stored at
-80C, and sectioned in a cryostat at -14C.  Sections are thaw-mounted onto
gelatin subbed or silylated (Onasco) slides.  After dessication overnight
(5C, with dryrite), slides are stored with a dessicant capsule at -80C
until assay.  Slides are brough to room temp and placed in preincubation
buffer (TrisCL, NaCl, etc) for 10 minutes, during which some parts will
fall off.  Anybody have similar experiences?  



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