| 主权项 |
1. A light-microscopy method for locating point objects in a sample arranged in an object space, the sample being imaged onto a detector by an imaging optical unit having a depth of field of predetermined axial extent along an optical axis in the object space, onto which the detector is imaged, wherein the point objects in the sample are located within the depth of field, wherein a first sample image generated by the imaging of the sample onto the detector is evaluated,
wherein, for locating a respective first point object in a direction of the optical axis, a parameter of a first light spot of one or more light spots of the first sample image representing the first point object is determined, and a first axial z position related to the first point object is assigned to the parameter based on predetermined association information, the method comprising: shifting the depth of field within which the point objects are located in the object space relative to the sample along the optical axis by a predetermined axial displacement, the predetermined axial displacement being smaller than the predetermined axial extent of the depth of field; imaging, by the imaging optical unit, the sample onto the detector with the axially shifted depth of field to generate at least a second sample image; determining second image z positions of the point objects in the second sample image based on the predetermined axial displacement; comparing first image z positions of the point objects in the first sample image to the second image z positions of the point objects in the second sample image; and generating, based on the comparing the first image z positions of the point objects in the first sample image to the second image z positions of the point objects in the second sample image, correction information so as to correct the first axial z positions related to the first point object based on the predetermined association information. |
