I am using a fluorescence microscope with DAPI filter, here are the specifications:
Excitation wavelength: 360/40 nm
Emission wavelength: 460/50 nm
Dichroic mirror wavelength: 400 nm
I want to label my cells with cyan fluorescent protein. I just want to know if our DAPI filter can detect the CFP. Thanks.
Probably not. A GFP filter would be more suitable. You can use a spectral viewer to see the emission spectrum of CFP and the overlap with common filters. Here the one from thermofisher: https://www.thermofisher.com/uk/en/home/life-science/cell-analysis/labeling-chemistry/fluorescence-spectraviewer.html#!/
Not ideal - although Cyan can be excited by the 405 nm laser line it is optimally excited at 485 nm. Could try using a standard FITC filter set (common to many epifluorescence scopes). See Chroma: https://www.chroma.com/products/sets/sp101-fitc
- Cyan GFP set/FITC Filter set
- Exciter 436/471nm
- Dichroic 455DLP/497LP nm
- Emitter 480/522 nm
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