Light-sheet microscope with parallelized 3D image acquisition

One or more tangible non-transitory computer-readable storage media storing computer-executable instructions for performing a computer process on a computing system, the computer process comprising:

• obtaining image data corresponding to a plurality of image planes of a sample, the image data representative of fluorescence emitted in response to illumination of portions of the sample using a plurality of beams of light, with different portions of the sample each illuminated in parallel by a different one of the plurality of beams of light and adjacent portions of the sample being illuminated by adjacent beams of light of the plurality of beams of light that are spatially separated with respect to each other;
• generating one or more images by reconstructing the plurality of image planes using virtual confocal slit apertures;
• and
• generating a three-dimensional reconstruction of the sample from the one or more images.

A system for image acquisition of a sample, the method system comprising:

• an illumination objective configured to illuminate the sample across a plurality of image planes using a plurality of beams of light, such that different portions of the sample are each illuminated in parallel by a different one of the plurality of beams of light, with adjacent portions of the sample being illuminated by adjacent beams of light of the plurality of beams of light that are spatially separated with respect to each other;
• a detection objective configured to detect fluorescence emitted in response to illumination of the plurality of imaging planes of the sample by the plurality of beams of light;
• and
• at least one processor configured to generate one or more images of the sample based on image data representative of the fluorescence emitted in response to the illumination of the plurality of imaging planes.