Posted by
Rosemary.White on
URL: http://confocal-microscopy-list.275.s1.nabble.com/5-color-imaging-6-color-would-be-even-better-tp7579662p7579668.html
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Dear Aromis,
5-colour imaging should be possible without bleed-through by sequential scanning on a regular confocal. Not sure how this is done with spinning disc instruments, but if possible, this is an option.
cheers,
Rosemary
Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia
T 61 2 6246 5475
F 61 2 6246 5334
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From: Confocal Microscopy List [
[hidden email]] On Behalf Of Aromis Storm [
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Sent: Saturday, 9 February 2013 1:13 a.m.
To:
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Subject: 5 color imaging (6 color would be even better!)
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Dear Everyone:
I have been trying to visualize a macromolecular structure with 4 color
imaging. I have been doing this with spinning disc microscopy as I need
complete sections of cells also to know the distribution of the structure
inside the cells in 3D. So far things are working properly BUT ...
We now would like to upgrade to 5 color imaging as we want to visualize
additional components of the structure. I checked the spinning disc setting
myself and I asked several senior users of spinning disc and got the opinion
that "5 color imaging with spinning disc without bleed-through is hardly
possible, if possible at all". The main reason being that it is not very
likely to set up emission filter combinations for 5 color imaging without
bleed-through. Spectral mixing is technically possible but not very
promising.
I also learned that 5 color imaging is possible with laser scanning
microscopy with which one can specifically define the emission band pass
range. But with laser scanning microscope it will cost me much more time to
collect enough data for statistical analysis.
So my questions would be: can anyone of you share your
experience/opinion/literatures on 5 color imaging? I also tried to look for
literatures describing multi-color (> 4 color) imaging (I think there must
be decent papers about it) but so far I have not found any. Maybe some of
you have some nice papers about this?
Thank you very much for your input.
Nice weekend!
Aromis