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 ?
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.