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Does anyone have information regarding the photostability of Alexa Fluor
555 and Cy3B in aqueous medium? I'd like to use one or the other in a
TIRF imaging scenario, and their photostability could definitely sway my
opinion one way or another. Any information would be appreciated, even
if you only have insight for one of the dyes, not necessarily in
comparison.
Cheers,
--
Samuel A. Connell
Imaging Facility Director
Department of Immunology
St. Jude Children's Research Hospital
332 North Lauderdale St., E7061
Memphis, TN 38105-2794
(901) 495-2536
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of George McNamara
Sent: Wednesday, April 11, 2007 1:23 PM
To: [log in to unmask]
Subject: Re: Alexa 532, 546 & 568 with 532 nm laser. .
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Hi Christophe,
How about evaluating Alexa Fluor 555 and Cy3B?
You can read about Alexa Fluor 555 at Invitrogen.com and Cy3B at GE
Heathcare (formerly Amersham Biosciences) website and various
articles about it, such as M. Cooper et al 2004 J Fluorescence
(http://www.springerlink.com/content/q3u3271467573733/).
Hopefully, some day GE or Alan Waggoner will commercialize Cy5B and
Cy3B-Cy5B.
best wishes,
George
At 09:59 AM 4/11/2007, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Dear all,
>
>The lab I'm in just bought the new Leica personnal/cheap confocal
>microsope, called the Leica SPE. The lasers are three diodes for green
>(blue 491 I think), red (green 532 nm) and far-red emitting dyes. We
>used to go to a SP2 confocal with a 546 nm red excitation line, or a
>widefield system where the filter is better used with a 568 nm emitting
dye.
>
>- Has anyone compared Alexa Fluor 532, 546 and 568 conjugated secondary
>antibodies ? 546 is rumored to bleach very fast, but we see it
bleaching
>much faster on our widefield setup (Cy3 filter) than with the 546 laser
>line of the SP2 confocal. I nevzer used Alexa 532 and I'm curious to
>hear comments before bying it.
>
>- As regards using the 532 line of the new SPE, do you think we should
>try Alexa 532 ? They do not seem to be commonly used. Furthermore, will
>the separation with Alexa 488 be good enough to really use it as a red
>dye in parallel with a green labeling (it's more yellow than red...) If
>not, do you know if we can catch Alexa 546 or Alexa 568 excitation with
>a 532 laser line ?
>
>
>Thank you for your help,
>
>
>--
>Christophe Leterrier
>
>Postdoc
>INSERM UMR641 Neurobiology of ionic channels
>IFR Jean Roche - Mediterranee University
>Marseille, France
George McNamara, Ph.D.
Sacramento, CA
916-429-7629
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