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Dear microscopists,
I have basic questions regarding 775nm STED-compatible mounting media and
DNA stains and I thought to ask the collective knowledge first before I
start reinventing the wheel:
1. Normally I use Mowiol+DABCO or 90% glycerol+DABCO (non shrinking
option). However, from previous discussions in this group it seems that
DABCO may not be the best all round anti-fade reagent to use. Therefore I
was wondering if there is also a commercial non-shrinking mounting medium
formulation that would be 775nm STED/far red dye compatible and have
improved anti-fade properties?
I though to try SlowFade Diamond but in older bulletins from Leica they
suggested to avoid it (not sure whether they meant Gold or Diamond
version). Has anyone actually tried SlowFade Diamond in 775 STED or can
suggest another commercial non-shrinking antifade alternative for 3D cell
imaging.
2. Due to their broad excitation/emission spectra I have avoided DAPI or
Hoechst stains in the samples. However, would there be a suitable
alternative DNA stain excitable by standard laser lines and compatible with
775 STED/far red dyes (not necessarily STEDable like SiR–Hoechst (PMID:
26423723))?
Thank you,
Jakub
--
Dr Jakub Chojnacki
IrsiCaixa AIDS Research Institute
Barcelona, Spain
[hidden email] <
https://twitter.com/IrsiCaixa>
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