Fluorescence
Fluorescence
Fluorescence
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Problem II: cross- emissionWhen emission spectra of two fluorophores overlap, emission from onechannel is recorded (and attributed) to the other channel.Example: ECFP and DsRed2 are excited by lasers (432/ 561nm), emissionis recorded by two PMTs after a beam splitter operating at 550nm.Solution: 1) Add extra filters – record DsRed2 starting from 590nm!2) Do not record both fluorophores at the same time!