Re: destroy alexa fluor fluorescence to allow restaining **Vendor Reply**

[Kilgore, Jason A.]

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** Vendor Reply **

Antibody stripping, as another commenter had mentioned, may be an option.  A low-pH glycine can be used.  Given that these are whole mounts, though, it may be more difficult to effectively strip.  

Other than antibody stripping, the only other way I'm familiar with is photobleaching.  The AF dyes are very photostable, but as long as you aren't using an antifade mounting medium, then you could blast the cells with intense visible wavelength light, such as with a mercury lamp, and it should photobleach the dyes most or all the way.

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions

Thermo Fisher Scientific
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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dale Moulding
Sent: Thursday, February 16, 2017 8:13 AM
To: [hidden email]
Subject: destroy alexa fluor fluorescence to allow restaining

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Dear all,
does anyone know of a way to irreversibly destroy alexa fluor fluorescence so a sample can be stained with a second set of dyes?
We do whole mount staining for 3 antigens with Alexa 488, 568 and 633. We would then like to re-image the same samples with cellmask and phalloidin.
Is there any way to kill alexa fluor fluorescence without destroying membranes and phalloidin binding?
Cheers
Dale