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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