Why is my yellow dye signal very weak in dSTORM imaging?

Dear all,

I’m suffering from a very weak signal of yelow dye, such as CF568 and JF549, in my dSTORM imaging. I used yellow dye-conjugated Halo tag or antibody to stain my samples. Althrough I can see a very bright and specific staining under TIRF or confocal microscope, the brightness of localizations under dSTORM is very weak and make it impossible to identify a single molecules. Does anyone has an experience on that? Any suggestions will be highly appreciated.

Attached is an example of my dSTORM image.

Thanks!

image

Can you tell us a little more about your imaging conditions? Illumination laser power? Microscope, and microscope objective (NA? oil? water)? Imaging buffer? Depth into the sample you are trying to image? Camera?

-Hazen

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