Methods and devices for imaging large intact tissue samples
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsRaju Tomer, Karl A. Deisseroth
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Abstract
Methods and devices for conducting high-speed, high-resolution imaging of large intact tissue samples are provided. Aspects of the methods include placing a sample in an optically homogenous sample manipulation component, performing a calibration procedure to align a light sheet and a detection focal plane at a plurality of locations within the sample, and performing an imaging procedure on the sample to collect an image from each location. The collected images are reconstructed to form a three-dimensional image of the sample. Devices for carrying out the steps of the methods are also provided.
Claims23
A microscope device comprising:
- an illumination beam path that comprises a light source;
- a detection beam path that comprises a camera;
- an optically homogenous sample manipulation component that comprises a sample chamber having a volume;
- a controller;
- a processor;
- and
- a computer-readable medium comprising instructions that, when executed by the processor, cause the controller to simultaneously:
- acquire a plurality of alignment parameters for a first location in a sample placed in the sample chamber by aligning a first light sheet plane with a first detection focal plane at the first location in the sample and thereby image the first location in the sample with the first aligned light sheet;
- and
- acquire a plurality of alignment parameters for a second location in the sample placed in the sample chamber by aligning a second light sheet plane with a second detection focal plane at the second location in the sample and thereby image the second location in the sample with the second aligned light sheet,
- wherein the imaged first location and imaged second location in the sample are used to generate a three dimensional image of the sample.
A method of imaging a sample using a microscope device, the method comprising:
- placing the sample in a sample chamber in an optically homogenous sample manipulation component;
- simultaneously:
- aligning a first light sheet with a first detection focal plane of a microscope device at a first location within the sample to determine an alignment parameter for the first location and thereby imaging by
- illuminating the first location with the first light sheet to align the first light sheet with the first detection focal plane and capturing a first image of the first location with the first light sheet;
- and
- aligning a second light sheet with a second detection focal plane of the microscope device at a second location within the sample to determine an alignment parameter for the second location and thereby imaging by illuminating the second location with the second light sheet to align the second light sheet with the second detection focal plane and capturing a second image of the second location with the second light sheet;
- and
- constructing a three-dimensional image of the sample using the first image and second image.
Dependencies are inferred from the claim text.