[Histonet] thick paraffin sections for immunostaining

[PALMER Jason (SVHM)]

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.palmer <@t> svhm.org.au



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