Re: Alexa 750
Posted by
Julio Vazquez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Alexa-750-tp1459204p1460983.html
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Hi Olivier,
I doubt the META detector can read emissions past 720-750 nm. Also, if you look at the excitation/emission spectra of Alexa 750, such as here:
you will see that with standard confocal lasers (488, 543/561/633), you will achieve an excitation efficiency of at most 10%. Finally, if you want to use a conventional detector, you would need an appropriate emission filter on your confocal, such as a longpass 730 or higher, which is probably not standard, and you probably would have rather poor detection efficiency, unless you have special IR PMTs...
One suggestion would be to use a dye in the red range that you can spectrally separate from Cy3, or maybe use something with a large stokes shift, such as a Qdot 605 or similar, that you can excite at around 450 or 488, and detect in the red channel, and therefore can be separated from Cy3 based on the different excitation efficiency. Something like Cy5-PE might work too (Excitation at 488, emission at 680)
--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024
On Nov 5, 2008, at 12:32 AM, Olivier Bardot wrote:
Hello
We are using DAPI, FITC, Cy3 and Cy5 for immunolabelling. We want to use a
fifth label and have found that Alexa750 seems to be a good possibility.
We are working on Zeiss LSM 510 Meta.
Does anyone have been using this dye?
Is it really possible to discriminate Cy3 from Alexa750?
I will appreciate any advices.
Thanks