I'm doing LSCM observations on fixed cells. I can not excite DAPI nor Hoechst dyes. Does anyone can suggest me a nuclear dye that is not emitting near the Alexa488 and 594, to prevent overlapping ?
Thanks
@Guillaume
Have you considered DRAQ5? Its excitation maximum is ~647 nm (red laser), its emission maximum ~665 nm. When intercalated with DNA, the excitation and emission maxima shift to ~681 nm and ~697 nm, respectively. It can be used with live or fixed cells. It is available from various vendors, so consider a vendor that you trust and who can give you good pricing. Hope this helps.
Hello Guillaume,
You probably need a far-red/near IR nuclear dye here; you may have a look at spectra plotting tools like SpectraViewer to make up your mind.
A possible setup would be TOTO-3 or DRAQ5 with a laser excitation at ca. 635 nm and a detection filter at ca. 690/50 nm.
http://www.thermofisher.com/ca/en/home/life-science/cell-analysis/labeling-chemistry/fluorescence-spectraviewer.html
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