[3dem] Neg-stain CLEM?

[jaime.llodra at ana.unibe.ch]

Dear Teige,


what is it that you want to image?. Dehydration destroys the fluorescence of fluorescent proteins.

Some fluorescent compounds like DAPI are destroyed by Uranyl Acetate.

If cells are attached to the grid under hydrated conditions then it can be possible to preserve their position after dehydration/negative staining and then perform CLEM.


Best,


Jaime Llodra



________________________________
Von: 3dem <3dem-bounces at ncmir.ucsd.edu> im Auftrag von Matthews-Palmer, Teige Rowan Seal <t.matthews-palmer14 at imperial.ac.uk>
Gesendet: Dienstag, 25. September 2018 15:12:12
An: 3dem at ncmir.ucsd.edu
Betreff: [3dem] Neg-stain CLEM?

Dear 3DEMers,

I’d be interested to hear from anyone who has tried, in the absence of a CLEM cryostage, to correlate objects between fluorescent light microscopy and negative-stain TEM.
Any success avoiding complete rearrangement / breaking on removing a coverslip from an EM grid?
Or any success getting a fluorescent signal out from underneath a dried coat of stain…?
Since this is for a bacterial suspension, negative stain is an obvious, but perhaps not the best fixation method.

Thanks for any info.
All the best
Teige

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