Hello,
I used the secondary fluorescent antibodies AlexaFluor488 and AlexaFluor555 together in my immunofluorescence experiments. Now I would like to evaluate the colocalization of the two marked targets in my photos. I have doubts about the possible cross-talk artifacts since I saw there is overlap in the spectra of this two dyes. What do you suggest to me? Would you use them together?
Thanks
Dear Chiara,
in my opinion it is possible to combine this dyes. Check your microscope setup carefully to be sure that there is no possibility for crosstalk.
Best regards
Franz
PS: If you don`t have the right microscope setup, buy another secondary Ab, it`s cheaper. :-)
Hi Chiara,
I completely agree with Franz. I have used a combination of Alexa488 and Alexa546 without problems. But it depends on the setup of the microscope.
With kind regards
Ronald
Confocal microscopy can do this. I wouldnt trust co-localized signals on a lesser microscope.
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