Method and light microscopy apparatus for producing an image of a sample

摘要

A sequence of individual images is acquired by imaging the sample through imaging optics onto an image sensor. For the acquisition of each individual image, the sample is provided with a marker pattern, in which individual markers can be imaged in the form of spatially separable light distributions through the imaging onto the image sensor. The centroid positions of the light distributions are determined and superimposed to form a complete image of the sample. According to the present invention, an image-drift-inducing temperature value (ΔT1, ΔT2, . . . , ΔTn) is measured during the acquisition of the sequence of individual images. A temperature-dependent drift value (ΔX1, ΔX2, . . . , ΔXn; ΔY1, ΔY2, . . . , ΔYn) is correlated to the image-drift-inducing temperature value (ΔT1, ΔT2, . . . , ΔTn) based on predetermined correlation data. The determined centroid positions are corrected based on the drift value (ΔX1, ΔX2, . . . , ΔXn; ΔY1, ΔY2, . . . , ΔYn).

主权项

1. A method for producing an image of a sample using a light microscopy apparatus including imaging optics and an image sensor, comprising the following steps:
acquiring raw data representing a sequence of individual images by sequentially imaging the sample through the imaging optics onto the image sensor; the sample being provided, for the acquisition of each individual image, with a marker pattern, in which individual markers can be imaged in the form of spatially separable light distributions through the imaging optics onto the image sensor; determining centroid positions of the light distributions, which represent the imaged markers in the respective individual images; and superimposing the centroid positions to form a complete image of the sample, wherein an image-drift-inducing temperature value (ΔT1, ΔT2, . . . , ΔTn) is measured during the acquisition of the raw data representing the sequence of individual images; a temperature-dependent drift value (ΔX1, ΔX2, . . . , ΔXn; ΔY1, ΔY2, . . . , ΔYn) is correlated to the image-drift-inducing temperature value (ΔT1, ΔT2, . . . , ΔTn) based on predetermined correlation data; and the determined centroid positions are corrected based on the drift value (ΔX1, ΔX2, . . . , ΔXn; ΔY1, ΔY2, . . . , ΔYn), and a drift-free said complete localization microscopy image of the sample is formed, based on the corrected centroid positions after acquiring the raw data representing the sequence of individual images.