Method and assembly for light sheet microscopic analysis of a sample

Abstract

The invention relates to a light sheet microscope which comprises an illumination device, which generates coherent illumination light for multiple illumination wavelengths, a beam forming module (8) for generating a light sheet from illumination light, an illumination objective (10) for illuminating a sample (1) with the light sheet, and a detection objective (12) for mapping the light, which is emitted by the sample, onto a flat detector, wherein the optical axis of the detection objective (12) includes an angle which differs by 0 DEG and 180 DEG from the optical axis of the illumination objective (10). In such a light sheet microscope, the beam forming module (8) comprises a phase-selective element (19) on which multiple selection regions (20) are arranged which are spatially separated from one another, wherein each selection region (20) is assigned to a specific illumination wavelength, and wherein a predefined phase distribution for the respective illumination wavelengths is applied to each selection region (20). In addition, the beam forming module (8) comprises means for sequential or simultaneous selection of the selection regions (20) according to the respective illumination wavelengths.

Description

technical field [0001] The invention relates to a method and a device for examining samples by means of a light sheet microscope, and mainly to the problem of analyzing samples labeled with various dyes. Background technique [0002] The study of biological samples has become more and more important in recent years, wherein the illumination of the sample is realized by means of a light sheet whose plane (light sheet plane) is at a non-zero angle to the optical axis of the detection (detection direction) intersect. Usually, the plane of the light sheet forms a non-zero, but usually not necessarily a right angle, with the detection direction (generally with respect to the optical axis of the detection objective). This research method is mainly used in fluorescence microscopy and is summarized under the term LSFM (Light Sheet Fluorescence Microscopy). An example is the method described in DE 10257423 A1 and in WO 2004 / 0535558 A1 based on the above-mentioned document and calle...

AI Technical Summary

Problems solved by technology

On the second hand, due to the specimen preparation and the size of the specimen chamber, the optical sheet is relatively thick and thus limited in the axial resolution that can be achieved

Third, specimen preparation is complex and not compatible with standard specimen preparation and standard assay holders typically for single-cell fluorescence microscopy

However, this has the disadvantage that at ~10ms exposure time, the optimization of the SLM has to be done 100 times per second for the new laser wavelength

With conventional nematic SLMs, this cannot be achieved, so ferroelectric SLMs with their very low optical efficiency and limited phase deviation must be used

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