Frozen sections for IHC
|From:||Gayle Callis <firstname.lastname@example.org>|
We do frozens exclusively for research IHC
snap freeze tissue, cut at desired thickness,air dry overnight, if in a
humid area use a dessicator with silica gel. Fix next day in acetone
75%/absolute alcohol 25% 5 min at RT, go directly to two changes of PBS,
Have used 4C acetone on air dried sections (30 min to 4 hours, even
overnight) but sections never looked as great as they do now. Remember
that some fixatives may work better for some antigens.
Have tried acetone/alcohol at 4C also, gave good morphology.
My murine CD4 is now out to 1:20,000 using this fixation protocol, so I
have not lost anything, but morphology is wonderful compared to acetone
We also do a glucose oxidase/glucose/sodium azide endogenous peroxidase
I can thank Barb Wright for this improved fixation method.
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
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