Hi Ian,

I think that the spectral imaging systems would be very reasonable for the
application of detecting 4, 5, or more fluorophores simultaneously. As has
been stated previously, in the Zeiss 510 META system the signals are
collected via a multi-channel detector that has 32 PMTs and the resulting
emission spectra can be unmixed. It has been my experience with this
technology that having spectrally well separated fluorophores and lasers are
much less of a concern. To get a better feel of how this might work for your
own goals, you might find the following article at
www.biosciencetechnology.com interesting, "Emission Fingerprinting Opens
Door To New Applications In Confocal Multifluorescence Imaging". You may
need to click on "Application Articles" in the main menu bar to get to the
appropriate link. This article shows 4 fluorescent proteins using
simultaneous imaging.

I actually will be trying to image 5 probes within the next few months on
the 510 META and will have a better feel of how this might work in real-life
experiments. I'll let you know how it goes.

Best Regards,

Kirk J. Czymmek, Ph.D.
Associate Professor
Department of Biological Sciences
Delaware Biotechnology Institute
University of Delaware
Newark, DE 19716




www.biosciencetechnology.com
[Kirk Czymmek]






>
>Hello Confocalists
>
>We are beginning to think about new microscopes and I'd appreciate any
>info on the following question:
>
>Is there a system out there which will allow simultaneous detection /
>imaging of four fluorophores emitting from blue to far red (eg AMCA /
>Cy2 / Cy3 / Cy5)? We do this sort of imaging routinely in wide-field and
>increasingly we need to it at confocal level. (Indeed if five channels
>were available we'd use them!)
>
>I know that the lasers are available (eg blue diode plus Kr/Ar or Ar
>plus red diode or whatever... I know that Leica and Zeiss have their
>spectral-detection systems (discussed here not long ago). From what I
>can figure out, it looks like BioRad has the lasers but only three
>detection channels; Zeiss seems to have the detection system but do they
>have the lasers? I got lost in Leica's website and gave up...(Sorry
>Leica!)
>
>Since the discussion a couple of months ago, has anyone come up with new
>info / ideas about the real value of the Leica and Zeiss approaches?
>From what has appeared on the list so far, I suspect there is a big gap
>between theory and practice here...
>
>Feel free to reply off list if appropriate.
>
>Thanks
>
>IAN
>
>
>
>--
>Professor Ian Gibbins
>Anatomy &Histology
>Flinders University of South Australia
>GPO Box 2100, Adelaide, SA 5001
>Australia
>
>Phone:  +61-8-8204 5271
>FAX:   +61-8-8277 0085
>Email:  [log in to unmask]