Scanning Luminescence Light Microscope with Gratings of Luminescence Inhibition Light and Further Light

摘要

A scanning luminescence light microscope for spatial high resolution imaging a structure marked with a luminescent marker comprises a light source for luminescence inhibition light and for further light; a light shaping and aligning device; and a detector registering luminescence light emitted by the luminescent marker. The device, by means of two optical gratings and an objective lens, forms two crossing line gratings of the luminescence inhibition light, and two crossing line gratings of the further light so that local intensity minima of an overall intensity distribution of the luminescence inhibition light are delimited in at least two directions, and that local intensity maxima or local intensity minima of an overall intensity distribution of the further light coincide with the local intensity minima of the luminescence inhibition light. Further, the device moves the overall intensity distributions of the further light and the luminescence inhibition light to scan the structure.

主权项

1. A scanning luminescence light microscope for spatial high resolution imaging a structure in a sample, the structure comprising a luminescent marker, the microscope comprising
a light source configured to provide luminescence inhibition light and further light differing from the luminescence inhibition light; a light shaping and aligning device including an objective lens and configured to form, from two non-coherent beams of the luminescence inhibition light, by means of two optical gratings, two pairs of coherent partial beams of the luminescence inhibition light and to focus and superimpose the two pairs of partial beams of the luminescence inhibition light by means of the objective so that the two pairs of partial beams of the luminescence inhibition light form two crossing line gratings of the luminescence inhibition light in the area of the sample, each of the two crossing line gratings of the luminescence inhibition light comprising a plurality of intensity minima delimited in one direction so that an intensity distribution of the luminescence inhibition light in the sample comprises a two-dimensional array of local intensity minima which are delimited in at least two directions,
wherein at least one beam of the further light enters the light shaping and aligning device together with one of the two beams of the luminescence inhibition light so that the light shaping and aligning device, from the at least one beam of the further light, by means of one of the optical gratings, forms two coherent partial of the further light and focuses and superimposes the partial beams of the further light by means of the objective so that the partial beams of the further light form a line grating of the further light in the area of the sample which comprises a plurality of intensity maxima delimited in one direction and intensity minima extending in parallel to the local intensity minima of one of the line gratings of the luminescence inhibition light;wherein the light shaping and aligning device is configured to align the line grating of the further light with regard to the one of the line gratings of the luminescence inhibition light in such a way that the local intensity maxima or the local intensity minima of the further light coincide with the intensity minima of the luminescence inhibition light;wherein the light shaping and aligning device is configured to move the intensity distributions of the further light and the luminescence inhibition light with regard to the sample to scan an area of interest of the sample with the intensity minima of the intensity distribution of the luminescence inhibition light; and a detector configured to register the luminescence light emitted out of the individual intensity minima of the intensity distribution of the luminescence inhibition light separately for each of the intensity minima and for each position of each of the intensity minima within the sample.