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I'd like to ask why are there no fluorescence microscopes that use a monochromator instead of filters?

If there are any, then I should change my question as, which ones use it and why are they so rare?

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  • $\begingroup$ Add Little bit of more data. $\endgroup$ Mar 10, 2015 at 13:50
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    $\begingroup$ Well, according to Cold Spring Harbor Protocols, xenon arc lamp/monochromator-based fluorescence microscopes are economical, performant, and rather common. $\endgroup$
    – MattDMo
    Mar 10, 2015 at 17:39
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    $\begingroup$ It is a very clear question about use of a specific piece in a laboratory equipment. It doesn't mean that a question is unclear, whenever you don't have the necessary background to answer it. I don't think each question should be asked by providing background information in such a way that, anybody related to biology should understand what is going on. Biology is a wide field and it is very unpractical, if this is what you are suggesting. And if not, I really can't see the point of putting this question on hold. $\endgroup$
    – Macond
    Mar 11, 2015 at 7:50

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Zeiss LSM780 uses essentially monochromator to spread all fluorescence across 34 detectors for multi-spectral acquisition. It is used to detect autofluorescence, unmix closely positioned emission spectra (unmix GFP and YFP for example) and to measure precise spectral peaks with ~1nm resolution.

Monochromator is useful when you need to transform frequency domain information into spatial domain. In fluorescence microscopy we usually collect light from wide bands of well-separated GFP-like proteins, so that filters make more sense. We usually don't detect 1-nm wide lines or plot spectra. LSM780 does allow all that, if you need to.

You can use spectral imaging, but for that you will have to either scan for very long time, or spread signal onto array of detectors.

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  • $\begingroup$ I'll add that signal is usually limiting in biological fluorescence microscopy, so it can be impractical or impossible to get good data in narrow wavelength bands. As a result most people use filters. And in turn this creates strong market pressure to design new fluorescent probes to match the spectra of existing probes and filters. $\endgroup$
    – TSwayne
    Apr 11, 2016 at 20:48
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    $\begingroup$ @TSwayne I think it is funnier: GFP and other proteins and dyes are/were designed to match laser lines (e.g. 488nm argon line). $\endgroup$ Apr 11, 2016 at 22:02

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