LIGHT-MICROSCOPIC METHOD OF LOCALIZATION MICROSCOPY FOR LOCALIZING POINT OBJECTS

摘要

A light-microscopic method of localization microscopy for localizing point objects in a sample arranged in an object space includes imaging, by an imaging optical unit having a depth of field range of predetermined axial z-extension along its optical axis in the object space, the sample onto a detector; localizing the point objects in the sample within the depth of field range in that, on the basis of a sample image, lateral x/y-positions of the point objects in a direction perpendicular to the optical axis are ascertained; displacing, in the object space relative to the sample the depth of field range within which the point objects are localized in the object space relative to the sample along the optical axis at least once by a predetermined axial z-travel distance; and imaging, by the imaging optical unit in the event of an axially displaced depth of field range.

主权项

1. A light-microscopic method of localization microscopy for localizing point objects in a sample disposed in an object space, the method comprising:
imaging, by an imaging optical unit having a depth of field range of predetermined axial z-extension along its optical axis in the object space, the sample onto a detector; localizing the point objects in the sample within the depth of field range in that, on the basis of a sample generated by imaging the sample onto the detector, lateral x/y-positions of the point objects in a direction perpendicular to the optical axis are ascertained; displacing, in the object space relative to the sample, the depth of field range within which the point objects are localized along the optical axis at least once by a predetermined axial z-travel distance which is smaller than the axial z-extension of the depth of field range; imaging, by the imaging optical unit in the event of an axially displaced depth of field range, the sample anew onto the detector to generate at least one further sample image; ascertaining the lateral x/y-positions of the point objects anew on the basis of the at least one further sample image; ascertaining lateral x/y-position deviations between the lateral x/y-positions of the same point objects in the sample image and the at least one further sample image; and generating correction information as a function of the lateral x/y-position deviations ascertained, using which information the lateral x/y-positions of the point objects which have been ascertained on the basis of at least one of the further sample images are corrected.