Re: Far-red filter set

Posted by Jennifer Waters on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Far-red-filter-set-tp592652p592670.html

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Here's my guess:
While the peak intensity of Cy5 is not visible, it starts emitting (if you look at the spectra) around 620nm which most of us can see.  So if there is a high enough concentration of probe, the small percentage of photons that emit from Cy5 in the lower wavelengths will be detectable.
Jennifer



On Fri, Mar 7, 2008 at 11:11 AM, Michael Herron <[hidden email]> wrote:
A nice paper on it here for wide field:  http://www.jhc.org/cgi/reprint/48/3/437

The Jackson website discusses Cy5 a bit here: http://www.jacksonimmuno.com/technical/f-cy3-5.asp

On Mar 7, 2008, at 9:27 AM, Martin Wessendorf wrote:

Search the CONFOCAL archive at

George Ring wrote:

SUPPOSEDLY the human eye can see up to 700nm, and women can see longer wavelengths than men.  That said, I agree with the others who responded - I haven't met anyone who can see it.  A camera, however, or confocal can detect far red.

I think that the human eye has been shown to be capable of detecting photons out beyond 1000 nm--it's just the probability of detection gets pretty low.

--Based on my own lab's experience, I agree with Peter's comment--I haven't met many people who can't see Cy5, for instance.  However, intensity is all-important.  Filled cells that've been stained with Cy5-streptavidin are easy to see with a 20x 0.7 NA lens by eye and occasionally visible even at 4x.  However, smaller structures with weaker labeling (e.g. terminals) are much harder to see and even with brightly labeled cells, the details can get lost.

Re: IR filters on CCD cameras--they indeed can be deadly when trying to image far-red fluorophores!

Martin
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--
Jennifer Waters, Ph.D.
Director, Nikon Imaging Center at Harvard Medical School