Posted by
Kevin Ryan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/5-color-imaging-6-color-would-be-even-better-tp7579662p7579663.html
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You might find Garini et al 1998 "Signal to Noise Analysis of Multiple Color Fluorescence Imaging Microscopy" (
http://itmhrt.ca/students/files/April2008/pdfs_for_journal_clubs/pdf4_Signal_to_noise_analysis-Cytometry_1999-Y_Garini.pdf), a useful reference.
Kevin Ryan
Media Cybernetics, Inc.
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Aromis Storm
Sent: Friday, February 08, 2013 9:13 AM
To:
[hidden email]
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