Hello!
I am currently conducting experiments in which I am staining myofibroblasts and myotubes. To distinguish cell types I would prefer to have a multicolour staining comprising of 4 fluorescent dyes, however our microscope has only three light cubes to detect fluorescent dyes (Texas Red, GFP and DAPI).
Is it possible to detect and distinguish two fluorescent dyes using the same light cube (e.g. Texas Red and Alexa fluor 568)? We have a EVOS FL Color Imaging Systems microscope.
Texas Red and AF568 have extremely similar excitation/emission spectra, and probably should not be used in combination if you have other options. On the EVOS the Texas Red light cube (AMEP4955) will pick up both Texas Red and AF568 with nearly equal efficiency.
Using the EVOS FL Color, there isn't a good way to get four channels out of 3 light cubes. You are limited to 3 colors with those light cube options. I would instead recommend the following four dyes/light cubes: DAPI, GFP, Cy3 (or Alexa Fluor 555), Cy5 (or Alexa Fluor 647). But this would require you to have Cy3 and Cy5 light cubes instead of a Texas Red light cube.
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