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Spectra of Commonly Used Fluorophores One of the most important concerns for biologists imaging immuno-stained samples is bleed-through, or detection of signal from two fluorophores at the same time. In almost all cases this can be avoided by properly adjusting the laser intensity used to excite the fluorophore as well as the filters used in the configuration. Also imaging sequentially instead of simulaneously can minimize bleed through. The following links and graphs are provided for your convenience to aid in identifying the optimal laser for fluorophore excitation and to ensure there is no chance that two fluorophores can be excited and detected in the same track of a configuration. A short example of how to use spectra is also included. Additional spectra, charts and references can be found online through several databases at: |
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GFP/YFP/CFP from http://dwb.unl.edu/Teacher/NSF/C08/C08Links/pps99.cryst.bbk.ac.uk/projects/gmocz/gfp.htm
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Cy3 dye from http://www.jacksonimmuno.com/technical/f-cy3-5.asp
Cy5 (see graph above under 543 laser line)
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