Patent ID: 6891613

Claim:
A flexible and freely programmable laser scanning microscope method for optical detection of quantities of different dyes in a specimen by detecting the wavelength-dependent behavior of an illuminated specimen in an laser scanning microscope using spectra which may contain heavily overlapping dye spectra from different dyes which are simultaneously examined, such as the emission behavior, absorption behavior, fluorescence, luminescence, phosphorescence, enzyme-active light emission, naturally occurring dye fluorescence from fluorescent proteins or enzyme-active fluorescence of the illuminated specimen, comprising: determining at least one spectral centroid position for different dye spectra detected by a spectral detector wherein a characteristic dye spectra is detected for at least one image point of the sample during a laser scan of the sample; and applying a characteristic spectral weighting function to the characteristic dye spectra that is detected to separate overlapping dye components and to calculate the intensity of each dye component per desired image point.