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Debbie,

        You don't explain the rationale for your rather
unusual choice of laser lines.  Our latest confocal (Nikon
C1) has 405nm, 488nm, 561nm which I find a tremendous
combination.  They are all solid-state so don't heat the
room and with the decent gap between 488 and 561 there
is a really good window for detecting FITC, GFP etc.
543 nm not only narrows that gap but it's a green HeNe
which is too gutless to be much use (we have two of these
so I know it's true!).  532 DPSS is great - powerful etc
but really cuts into your 'window' for FITC.  If I wanted to
add a red laser (which I don't see much need for) I'd go
for a diode at 647 which again leaves you a decent gap
beyond the 561.  Much more useful than 633 HeNe.

Guy

Debbie Hardie wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Fellow confocalists!
>
> Has anyone any experience of the relatively new 561 'Lime' laser? We are choosing a new confocal system and want as many colours as possible. Along with 405 and 488 excited dyes its likely we'll use three red dyes TRITC or Cy3, Texas Red, and 633 or Cy5.
> We have to chose between the 543, 594 and 633 combination or the 561, 594 and 633 combination.
> On its own the 561 is more powerful (10mW as opposed to 1.5mW for the 543), but the question remains which is the best combination? I'm inclined to ignore the extra power of the 561 and go for the 543 as it gives better separation.
> Has anyone tried the 561?
> Thanks
> Debbie
>
> Debbie Hardie
> Research Fellow
> MRC Centre for Immune Regulation
> Institute of Biomedical Research Floor 4
> University of Birmingham
> Birmingham
> B15 2TT
>
> 0121 414 6970

--
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Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
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Phone +61 2 9351 3176     Fax +61 2 9351 7682
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