Method for correcting background signals in captured measurement values of analog detectors

A method for operating a fluorescence microscope to correct background signals in captured measurement values of an analog detector, the method comprising:

• guiding radiation emitted from a sample under fluorescence excitation along a detection beam path through optical elements configured to guide and shape the emitted radiation and to direct the emitted radiation onto detector elements of the analog detector;
• receiving, by an analog-to-digital converter, electrical signals corresponding to measurement values captured by the analog detector and digitizing the measurement values:
• and
• receiving, by an evaluation unit, the digitized measurement values from the analog-to-digital converter:
• analyzing, by the evaluation unit, measurement values of an object captured over a reference time period during the fluorescence microscopy measurement;
• determining, by the evaluation unit, characteristic values of captured background signals;
• detecting and correcting, by the evaluation unit dark signals of the analog detector from the background signals, the dark signals comprising output signals of the detector generated in absence of incident photons under the fluorescence excitation;
• determining, by the evaluation unit, a threshold value based on of at least one of the characteristic values and by applying a calculation specification;
• applying, by the evaluation unit, the threshold value to captured measurement values of the analog detector;
• and
• filtering, by the evaluation unit, the captured measurement values such that only those measurement values which are greater than or equal to the threshold value are used for a subsequent signal evaluation,
• wherein the analog detector is an array detector, and
• wherein during the fluorescence microscopy measurement, the detection and correction of dark signals by the evaluation unit is performed separately for each detector element of the array detector for fluorescence correlation spectroscopy analysis.