The DAPI stain turns yellow after 24+ hrs and I loose most Alexa 488/594 signal. Product has not expired, standard double label immuno protocol I have used for 20+ years. Had this problem in the past but used a new bottle of VectaShield and the color shift of the DAPI when away. Sometimes half of the mouse brain section is OK and half is yellow/orange in the DAPI channel. My immunostaining should last for months at 4C in the dark. Can't afford to have my experiments ruined! Does anyone have a better, more consistent mounting media to try?
I'm with Thermo Fisher Scientific technical support. I recommend using our ProLong Glass with NucBlue (Hoechst 33342). ProLong Glass solidifies for archiving long term (though I still recommend storing the slide at -20C after a couple days of hardening at RT), doesn't quench Alexa Fluor dyes, doesn't yellow, and has superior ability to slow photobleaching. It is also made for tissue sections as it has a refractive index that matches glass (thus the name) allowing for deeper imaging of thick sections and refractive index matching with immersion oil and glass lenses. The 2 mL trial size is catalog P36983.
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