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Michael,

We also have observed that Alexa 647 phalloidin can be excited by 488
nm with emission around 525 nm.  With 633 nm excitation, the emission
is stronger and the peak is around 675 nm.  This was measured with
the Zeiss 510 Meta.  The Alexa 633 phalloidin seems cleaner; it
excites well at 633 nm and emits around 640 nm.  We get good results
using the 633 phalloidin. Regards, Carolyn Smith

PS. If this message repeats, I apologize but it seemed to bounce back
to me the first time I sent it.


>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>We're having an odd problem.
>
>We're staining COS cells with Alexa 647 phalloidin (Mol. Probes A-22282 lot
>41B1-1).  The f-actin is staining fine and looks great when we excite at
>633 nm and detect through the Leica AOBS or normal Cy5 filter set.
>
>However, when we excite at 488 nm, we're seeing the f-actin staining
>appearing in the green channel too.  We checked on different microscopes.
>
>Has anybody else seen something as weird as this?  We suspect
>contamination, but don't know where it would have come from unless it is an
>isomer or something from the manufacturing process.
>
>One of our controls is the Alexa-phalloidin staining alone without any
>antibodies, so we know it's not some weird antibody binding artifact.
>
>Any help appreciated.
>
>Thanks.
>
>____________________________________________________________________________
>Michael Cammer   Analytical Imaging Facility   Albert Einstein Coll. of Med.
>Jack & Pearl Resnick Campus      1300 Morris Park Ave.     Bronx, NY  10461
>(718) 430-2890       Fax:  430-8996      URL:  http://www.aecom.yu.edu/aif/

Carolyn Smith
Manager, Light Imaging Facility
NINDS/NIH
Building 36 Room 3B20
Bethesda MD 20892

Telephone: 301-496-8893
Email [log in to unmask]