Light sheet microscope and method for light sheet microscopy

A method for light sheet microscopy, comprising:

• illuminating a sample with a light sheet comprising a light sheet plane, wherein the light sheet plane is arranged substantially perpendicularly to an optical imaging axis and the light sheet has an illumination width extending perpendicular to the light sheet plane;
• setting the light sheet plane sequentially to different locations of the light sheet plane of the light sheet in the sample;
• recording an image of the sample for each of the different locations of the light sheet plane, thereby producing a stack comprising of a plurality of images of the sample, with each image in the stack corresponding to a single one of the different locations of the light sheet plane,
• wherein a spacing, measured along the optical imaging axis, between adjacent locations of the different locations of the light sheet plane is greater than or equal to half the illumination width, but not greater than the illumination width;
• and
• computing intermediate images, each of the intermediate images corresponding to a virtual additional location of the light sheet plane in the sample.

A light sheet microscope, comprising:

• a sample volume, comprising a sample stage for a sample carrier with a sample;
• a detector that images the sample volume along an optical imaging axis;
• an illumination source that illuminates the sample with a light sheet, comprising a light sheet plane that is adjustable in the sample volume, wherein the light sheet plane is arranged substantially perpendicularly to the optical imaging axis, and wherein the light sheet has an illumination width extending perpendicular to the light sheet plane;
• and
• a processor, which is connected to the detector, and is connected to the illumination source or to the sample stage or to both,
• wherein the processor is configured to:
• control the illumination source or the sample stage, or both, to set a location of the light sheet plane sequentially to different locations of the light sheet plane of the light sheet in the sample volume,
• control the detector to record an image of the sample volume for each of the different locations of the light sheet plane, whereby a stack of a plurality of images of the sample volume is produced, with each image in the stack corresponding to a single one of the different locations of the light sheet plane,
• control the illumination source or the sample stage, or both, such that for the plurality of images recorded a spacing, which is measured along the optical imaging axis, between adjacent locations of the different locations of the light sheet plane is greater than or equal to half the illumination width, but not greater than the illumination width, and
• compute intermediate images for the stack, each of the intermediate images corresponding to a virtual additional location of the light sheet plane in the sample volume.