We would like to do immunostaining on thick FFPE sections for neuronal markers
such as tyrosine hydroxylase so that we may trace neurons contained in
non-brain tissue. Section thickness will probably be in the order of 40-50
microns, though perhaps thicker too. I have searched histonet archives but
have only found references to vibratome sections of brain tissue, wholemount
embryos and free-floating sections that I assume were cut on a cryostat.
Antibody penetration has been achieved using DMSO, ethanol extraction, acetone,
triton-x 100 etc, but I was wondering if anybody had managed to use any of
these methods successfully on thick paraffin sections of non-brain tissue - our
tissue is predominantly connective / adipose in nature. It is possible that we
could do vibratome sections if we had to, but the tissues have already been
collected, processed and paraffin blocked and would rather not have to do
further experiments and tissue collection.
I did also note in the archives that in regular paraffin sections, antibody
penetration and thus labelling with standard methods is only thought to be of
the order of 2 microns.
Many thanks,
Jason
Jason Palmer
Bernard O'Brien Institute of Microsurgery
42 Fitzroy St, Fitzroy Victoria 3065
Australia
tel +61 3 9288 4018
fax +61 3 9416 0926
email: jason.pal...@svhm.org.au
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