AW: [Histonet] Immunofluorescence on FFPE skin

[Gudrun Lang]

HI Erin,
we tried to do the usual IHC-protocol for immunoglobulins on fixed
skin-biopsies for Pemphigus. (on Ventana Benchmark, with HIER in high pH
retrieval buffer). It gave very, very overstained sections. We didn't
continue this project, because usual IF is easier to "fix" , has worked "for
ever" and the results are typically described in pathology-literature.

But I think it could work, if you make a really high dilution of the
antibodies. And in mind of the brown melanin-pigment, a red chromogen would
give a better contrast.
In my opinion microwave plays no role - it just has to be HIER.

Gudrun


-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Martin,
Erin
Gesendet: Montag, 18. März 2013 19:28
An: histonet
Betreff: [Histonet] Immunofluorescence on FFPE skin

Hello all,



My pathologist gave me a copy of "Immunofluorescence with Dual Microwave
Retrieval of Paraffin-Embedded Sections in the Assessment of Human Renal
Biopsy Specimens" from the American Journal of Clinical Pathology.  He said
that the same principle should work on skin and he would like to be able to
do IF on fixed tissue in addition to our usual cryostat sections.  Has
anyone else read the paper who might be willing to give me some basic advice
to try working it out?  Is a microwave necessary (paper's method uses 2
different wattage settings) or is there a way to use HIER in waterbath or
pressure cooker?



Thanks in advance,

Erin



Erin Martin, Histology Supervisor
UCSF  Dermatopathology Service
415-353-7248

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