Laser scanning microscope and method for operation thereof

摘要

Laser scanning microscope and method for the operation thereof having at least two detection channels which has at least one beamsplitter with a splitting of the sample light deviating from the 50:50 split and/or, with 50:50 split in the detection channels, has detectors with differently adjusted gain, or in at least one detection channel with equal light splitting has an additional light attenuator.

主权项

1. A method for operating a laser scanning microscope, the method comprising:
carrying out a two-dimensional horizontal image scanning in a plurality of focus positions or height positions; detecting, accordingly in a channel of each of at least two detectors, the sample light; recording three-dimensional image stacks simultaneously with the at least two detectors; splitting the sample light unequally between the at least two detectors with a color neutral beamsplitter; and calculating, for individual picture points and an associated height position, a measured intensity value of each of the two detector channels by picture point; wherein, when each of the two intensity values lies within a threshold region of the respective detector channel, a sample image is generated from the two calculated intensity values in that an average is formed between the two intensity values; wherein, when only one of the two intensity values lies within a threshold region of the respective detector channel, a sample image is generated from only one of the two intensity values; and wherein, when neither of the two intensity values lies within a threshold region of the respective detector channel, a sample image is generated using neither of the two intensity values.