[Histonet] immunostaining with Technovit 8100 / glycol methacrylate

[PALMER Jason (SVHM)]

Hi Histonetters.

I'm trying to immunostain some sections of rat tissue embedded in Technovit 8100 (a glycol methacrylate) with an ED1 antibody (Serotec rat anti mouse), and have had no success to this point. Have tried various antibody concentrations and antigen retrievals including pronase, trypsin (which we use successfully for immunostaining with the same Ab on paraffin-embedded tissues) and high temperature citric acid buffer retrieval. Our samples are paraformaldehyde fixed overnight, and we use a labelled streptavidin-biotin system. The technical information that comes with the product suggests it is suitable for immunohistochemistry (and this is why we purchased it, as well as its water miscibility and lack of requirement for heat or solvents during processing / embedding) but I been unable to find any reference to its use in this way in the scientific literature, or on the net in general, so far. Can anybody give me any suggestions at all as what I might try, or how I might be able to find useful information on immunos with glycol methacrylate in general or Technovit 8100 in particular? I did see recently an archival Histonet posting on "etching" of glycol methacrylate sections using a cured (ie aged) NaOH solution - does anybody have an opinion on this?

Many thanks,

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: palmerj <@t> svhm.org.au


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