| 摘要 |
The invention relates to a light-microscopy method for locating point object in a sample (76), wherein the sample (76) is imaged onto a detector (14, 56) by means of an imaging optical unit (70) and the point objects contained in the sample (76) are located within the depth of field (78) by evaluating a first sample image produced by the imaging of the sample (76) on the detector (14, 56). In order to locate the particular point object in the direction of the optical axis (O), a characteristic value (d) of a light spot (38, 40, 42, 44, 60) of the first sample image representing the point object is determined and an axial z position related to the point object is associated with said characteristic value (d) in dependence on predetermined association information. The depth of field (78) is shifted in relation to the sample (76) along the optical axis (O) by a predetermined axial displacement that is less than the axial extent (t) of the depth of field (78). While the axially depth of field (78) is shifted, the sample (76) is imaged onto the detector again by means of the imaging optical unit (70) and at least one second sample image is produced. In the second sample image, the z positions of the point objects are determined in dependence on the predetermined axial displacement. The z positions of the point objects determined in the first sample image are compared with the z positions of the same point objects determined in the second sample image. In dependence on this comparison, correction information is produced, on the basis of which the z positions of the point objects determined in dependence on the association information are corrected. |
