Dear Johan,
Perhaps this laser has a weak 568 nm output, or PMT1 is ailing. Have
you measured the output at 568 nm with a laser power meter?
Without a power meter, you might improvise by comparing intensity of
emissions from a dye exciting at both 488 and 568 or 568 and 647 nm. In
the latter, Molecular Probes' Deep Red fluorescent nissl stain emits
nearly equally when excited at 568 (605/32 filter, PMT1) or 647 (680/40,
PMT3).
If you remove filter block 2 is PMT1 noisy with a green emission, in
comparison to PMT2?
Regards,
Glen
Johan Wassélius wrote:
>
> Dear fellow confocalists!
>
> I have encountered a problem with our BioRad 1024, and since neither myself
> our technician can solve it, I turn to your expertise for help, as I have
> done before with great results.
>
> Our BioRad 1024 has since the laser was last replaced been very weak in the
> red (Texas Red) channel. The staining looks exactly the same as before when
> I look at it through the oculars, but I have to maximize the laser power
> and the settings (iris and almost the gain to), to get anything on the
> screen and by doing so I obviously get an image that is so very noisy that
> it is completely useless.
>
> In the FITC or cy5 channels I can use the same, or at least very similar,
> settings as earlier, with the same image quality as earlier, so the problem
> is entirely, or almost entirely, focused to the TR-channel.
>
> Has anyone of you out there experienced this problem? Suggestions on how to
> solve it will be greatly appreciated!
>
> Cheers,
>
> Johan
>
> ~~~~~~~~~~~~~~~~~~~~
> Johan Wassélius
> Wallenberg Retina Center (WRC)
> BMC, B 13, Klinikgatan 26
> Lund University Hospital
> S-221 84 Lund, Sweden
> Phone: +46-46-222 07 65
> Fax: +46-46-222 07 74
> Home phone: +46-46-211 99 93
> Cell phone: +46-0703-750593
> e-mail: [log in to unmask]
> ~~~~~~~~~~~~~~~~~~~~
--
Glen MacDonald
Microscopy and Imaging Facility
University of Washington Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923
[log in to unmask]
(206) 616-4156 (206) 616-1828 fax
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