Method of using a high resolution laser scanning microscope and high resolution laser scanning microscope

摘要

A high resolution laser scanning microscope has beam shaping elements configured to shape a beam of fluorescence inhibiting light which is directed into a back aperture of an objective connected to form an intensity minimum delimited by intensity maxima of the fluorescence inhibiting light in a focus of the objective. A plurality of optical elements including the objective and the beam shaping elements are arranged in a beam path of the beam to the focus. Using the microscope includes removing or exchanging or altering or adding at least one of the optical elements arranged in the beam path of the beam of fluorescence inhibiting light, and compensating a variation of polarization varying properties of the plurality of the optical elements, that is caused by removing or exchanging or altering or adding the at least one optical element, by adapting the beam shaping elements to the variation.