We are planning to do a pre-embedding immunoreaction for EM. Although there are many alternatives we will try the DAB reaction on HRP-conjugated Ab, followed by silver precipitation (Gallyas, 1982) and gold substitution: GSSP-protocol.
One of the key chemicals of the GSSP-procedure is gold chloride (AuCl3). We don’t have that anymore. What we do have is the Nanoprobe GoldEnhance for EM plus (GEEM+). Do you know if this GEEM-solution can be used to replace the silver-precipitate?
Thanks for your time
Dear Duko,
in theory you can use the Nanoprobe GoldEnhance as it works on a similar principle.
Regards.
Adalberto
I've never tried this one but as above in principle it should work! Only way to find out is to try it! You could also try tetramethylbenzidine instead of DAB! Said to be less toxic than DAB (little evidence) and generates crystals that shine brightly in polarised light illumination!
Dear Duko, I do not forsee any problems. The gold enhancement kits are working along the same principles.
Yes, it will work.
I also have made a ruthenium substrate and silver enhancer to go with it.
One can do both fluorescence and EM with ruthenium/silver.
Helaas heb ik hier geen ervaring mee (ik werkte vnl. met goud-partikels, post embedding), maar zo te lezen heeft u reeds enige goede antwoorden gekregen. Vriendelijke groet
There should not be any problem to use the Nanoprobe GoldEnhance to replace the silver precipitate
Thanks for all the encouraging answers. I will certainly try this out now and let you know!
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