[Histonet] Autofluorescence Issue

[Shawn Polson]

Hello,

We are running into a problem with what appears to be fairly broad 
spectrum autofluorescence (showing up on DAPI, FITC, and TRITC filter 
sets) emnating from tissues in our samples.  The tissues in question are 
acroporid coral fixed in Z-fix.  Samples were all processed by enrobing 
in agarose, decalcifying with EDTA, and embedding in paraffin.

Our intent is to use 16S rRNA FISH to identify bacteria within these 
tissue sections.  Does anyone know of an effective way to subtract out 
such autofluorescence either chemically (without damaging FISH targets) 
or by use of some virtual subtraction technique.

Thank you,
Shawn

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Shawn W. Polson, M.S.

Molecular Biologist, Coral Health Program
Center for Coastal Environmental Health and Biomolecular Research
NOAA National Ocean Service
Contractor, JHT Incorporated
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Ph.D. Candidate
Marine Biomedicine and Environmental Sciences Center
Molecular and Cellular Biology and Pathobiology Program
Medical University of South Carolina

Mailing Address:
Hollings Marine Laboratory
331 Fort Johnson Rd
Charleston, SC 29412
Office (843)762-8840     Lab (843)762-8959
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