[Histonet] Immunostaining on plastic embedded tissue

Herrick, James L. Herrick.James <@t> mayo.edu
Thu Dec 20 11:27:33 CST 2007


I am trying immunostaining (for actin and Von Willebrand Factor) on MMA
resin embedded arterial tissue with little success at this point. I
sectioned at  5um and 10 um thicknesses and deplastifyed with a couple
of 3 minute rinses of 2-Methoxyethyl acetate, followed by a quick
(approx. 20-30 sec.) rinse in chloroform. My sections have been sitting
out at room temp (following deplastification) for about 2 weeks before I
begin the immuno staining -- should they be stored in the refrigerator
or freezer? The sections look great and the plastic appears to be
completely dissolved, but I am thinking I may need to treat longer.
Also, I may try some type of antigen retrieval. Does anyone have
experience with this type of staining and if so, could you please give
me some tips on what I may need to change. Thanks for your help.


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