Method for localizing and tracking emitters in a sample
AssigneeAbberior Instruments GmbH
InventorMatthias Reuss
Life of this patent
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Abstract
The present disclosure relates to a method for localizing or tracking individual emitters in a sample according to the MINFLUX principle, wherein the sample is illuminated at several illumination positions in a close range around the emitter from different illumination directions with modulation light which comprises a linearly or flatly extended intensity minimum. A position of the emitter in the sample is calculated from the emissions of the emitter detected at the illumination positions. The invention further relates to a light microscope for carrying out the method.
Claims22
A method for localizing or tracking an individual emitter in a sample, wherein the sample is illuminated through an objective lens
- with excitation light, which excites the individual emitter to emit light, and
- with modulation light, which attenuates, amplifies, or excites the light emission of the emitter,
- wherein the modulation light forms an intensity distribution in the sample with an intensity minimum extended along a minimum line or a minimum area, in particular with a zero line or a zero area, the method comprising:
- detecting light emissions of the emitter for several illumination positions of the modulation light in a close range around the emitter, and
- calculating a position of the emitter in the sample from the emissions detected for the illumination positions,
- wherein the sample is illuminated with the modulation light in different illumination directions during the detection of the light emissions, wherein the light emissions are assigned or assignable to the different illumination directions.
A light microscope for localizing or tracking an individual emitter in a sample comprising:
- a light source for excitation light, which is adapted to excite the individual emitter in the sample to emit light;
- a light source for modulation light which is adapted to attenuate, amplify or excite the light emission of the emitter in the sample;
- an objective lens which is configured to illuminate the sample with the excitation light and the modulation light;
- optical means which are configured to form an intensity distribution of the modulation light in the sample in such a way that the intensity distribution comprises an intensity minimum extended along a minimum line or a minimum area, in particular a zero line or a zero area;
- a scanner which is configured to position the intensity distribution of the modulation light in the sample at different illumination positions in a close range around the emitter;
- and
- a detection device which is configured to detect light emissions of the emitter for the illumination positions,
- wherein the light microscope comprises means which are configured to illuminate the sample with the modulation light from different illumination directions, and a computing unit which is configured to calculate a position of the emitter in the sample from the light emissions detected during the illumination of the sample with the modulation light in different illumination directions.
Dependencies are inferred from the claim text.