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COMMERCIAL RESPONSE

Hi Dale,

I agree with Kelly that you can do this with 5 colors relatively
easily *(although
commercial responses should be labeled as such).*

Below is a link to a slide deck on our website outlining the imaging scheme
and filter sets to best separate these.  BD Biosciences also offers the
BV480 (like Aqua or CFP, but much, much brighter) which allows for distinct
excitation and emission ranges for all 5 fluors.

https://www.chroma.com/sites/ <goog_1687719810>default/files/5-CHANNEL%
<goog_1687719810>20FLUORESCENCE%20IMAGING% <goog_1687719810>
20SIMPLIFIED%20-%20Reliable% <goog_1687719810>20Multiplexing%20for%20the%
<goog_1687719810>20Non-Specialist_New.pdf

Chroma does not profit from sales of any of the fluorophores, only from the
filters.  We also offer two different 5-band filter sets to accommodate
this:

https://www.chroma.com/products/sets/89903-et-bv421-bv480-af488-af568-af647-quinta-band-set
https://www.chroma.com/products/sets/89904-et-405-445-514-561-640nm-laser-quinta-band-set

I was initially skeptical that we could achieve good separation between
BV480 and AF488, but with the right filter sets (specified in the slide
deck) you can indeed.

Good luck,
Jeff



*Jeff Carmichael*

*Technical and Product Marketing Manager*

*[log in to unmask] <[log in to unmask]>*Chroma Technology Corp.

*an employee owned company*
*10 Imtec Lane*
*Bellows Falls, VT  05301*
*802-428-2528 Office*
*802-428-2528 Fax**800-824-7662 Toll Free*

On Thu, Feb 16, 2017 at 11:12 AM, Dale Moulding <[log in to unmask]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> 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
>