Simultaneous orthogonal light sheet microscopy and computed optical tomography
AssigneeCalifornia Institute of Technology
InventorsJohn M. Choi, Thai V. Truong, David S. Koos
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Abstract
An apparatus for and method of performing orthogonal light sheet microscopy (OLM) and computer optical tomography (COT) simultaneously in a single device are provided. The dual-mode imaging microscope allows for the use of both OLM and COT in a single instrument. This dual-mode device will allow researchers to have access to both types of microscopy, allowing access to the widest possible selection of samples, and improved imaging results. In addition, the device will reduce the high costs and space requirements associated with owning two different imagers (OLM and COT).
Claims42
A dual-mode imaging microscope comprising:
- at least one excitation source, said excitation source being capable of producing an excitation beam, and being disposed such that the excitation is directed along at least one excitation beam path;
- a sample holder in optical communication with said excitation source such that a sample contained therein is excited by said excitation beam;
- an excitation optic capable of producing from the excitation beam a substantially two-dimensional sample excitation region having an effectively uniform excitation intensity which extends in the direction of an excitation axis of the excitation beam path and transversely thereto and intersects with at least a portion of the sample holder along a pre-determined path;
- a tomography detector configured to detect an excitation generated signal contrast from the sample holder, the tomography detector being disposed such that the axis of detection of at least one tomography detector is substantially collinear to the excitation beam;
- a light sheet microscope detector configured to detect an excitation generated signal contrast from the sample excitation region, the light sheet microscope detector being disposed such that the axis of detection of at least one light sheet detector is substantially orthogonal to the sample excitation region;
- wherein the angular orientation of the excitation region relative to the sample holder may be varied about the axis of the sample holder, and wherein the position of the excitation region relative to the sample holder may be varied along the light sheet detector axis;
- and
- wherein the axis about which the angular orientation may be varied is not parallel to the excitation beam path.
A method of imaging an object using a dual-mode imaging microscope comprising:
- producing an excitation beam, and directing said excitation beam along an excitation beam path;
- placing a sample within the optical path of said excitation beam to generate at least one signal contrast;
- producing from said excitation beam a substantially two-dimensional sample excitation region having an effectively uniform excitation intensity which extends in the direction of an excitation axis of the excitation beam path and transversely thereto and intersects with at least a portion of the sample;
- detecting said signal contrast along a tomography detection axis that is substantially collinear to the excitation beam;
- detecting said signal contrast along a light sheet detection axis that is substantially orthogonal to the sample excitation region;
- and
- varying the angular orientation of the excitation region relative to the sample about the axis of the sample and varying the position of the excitation region relative to the sample along the light sheet detector axis to obtain data along a specific angular and positional axis of the sample, wherein the axis about which the angular orientation may be varied is not parallel to the excitation beam path;
- and
- repeating the steps of producing, detecting and varying until sufficient angular and positional data has been obtained.
Dependencies are inferred from the claim text.