[Histonet] beginner question about methacrylate resins and nucleic acid ISH

From:Sheri L Simmons

I'm a newcomer to the list and the world of histochemistry in general. I'm 
hoping that someone can help me with a few questions relating to 
targeting bacterial RNA in embedded thick sections (very thick- about 
a mm) with fluorescent methods.
The goal is to detect species-specific oligonucleotide probes to 
the bacterial 16s rRNA with indirect CARD-FISH. Searching the list 
archives, it seems that although glycol methacrylate (GMA) and methyl 
methacrylate (MMA) resins are both used for 
histochemistry, MMA is generally preferred because it can be removed from 
sections with acetone. Is this your experience? Do you know of any direct 
comparisons of GMA and MMA for nucleic acid detection with in situ 
hybridization? Are there any special concerns in terms of 
the resin type or protocol relating to the use of nucleic acid probes rather than antibodies? 
Finally, which of the many MMA kits out there would you recommend? I've 
seen references to Technovit 9100, but this is significantly more 
expensive than a generic Methyl Methacrylate/Butyl Methacrylate kit from 
Electron Microscopy Sciences. Is there a reason to prefer one over the 


Sheri Simmons
Geomicrobiology Group
MS #52, WHOI, Woods Hole MA 02543

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