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** Vendor reply **

Polystyrene microspheres, such as the TetraSpecks, will swell in the presence of certain solvents.  So you'll want to use an aqueous-based mounting media, preferably one that cures (such as ProLong products or Fluoromount-G) instead of  using an organic solvent-based one, like Cytoseal-60.  When dye-labeled microspheres like these swell, the dye is free to diffuse out of the polystyrene matrix.

Cheers,

Jason


Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Thermo Fisher Scientific
 
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From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Jean Ross
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Subject: Tetraspeck beads for PSFs

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Hi Everyone,

I have been preparing 0.1um Tetraspeck bead slides to use when measuring point spread functions on our Zeiss Elyra PS1 super resolution microscope
and Zeiss LSM880 confocal with Airyscan.   The problem is that over time
the beads appear to swell to about 3 times in size and lose much of their fluorescence.  I have tried many different mounting medias (hardening and
nonhardening) all with the same result.  Has anyone else experienced this problem?  Any suggestions to solve the problem would be appreciated.

Thanks,
Jean

--
Jean Ross
Delaware Biotechnology Institute, BioImaging Center University of Delaware
15 Innovation Way
Suite 117
Newark, DE  19711
Phone:  (302)831-0620
Fax:  (302)831-4841