Choosing fluorochromes and fluorochrome combinations

Single labeling

• signal strength
• background
• suitable excitation sources (Hg lines, laser lines)
• suitable detection filter combinations (also dependent on excitation source)

Dual or triple labeling

• relative signal strength
• possible FRET (fluorescence resonance energy transfer), if emission spectrum of a fluorochrome overlaps with excitation spectrum of another fluorochrome in use and, in addition, the distance between both is <10 nm)