US 11,454,795 B2 · Surface sensing in optical microscopy and automated sample scanning systems

Abstract

The disclosure features methods and systems that include positioning a surface of a coverslip overlying a sample relative to an object plane of a microscope system, projecting a two-dimensional pattern of light onto the surface, where a focal plane of the two-dimensional pattern at a position of the surface is rotated by an angle β relative to the object plane, obtaining a two-dimensional image of the pattern of light reflected from the surface using a detector that includes an imaging sensor oriented perpendicular to a direction of propagation of the reflected pattern of light at the sensor, analyzing the image to determine a line of best focus of the pattern within the image, determining an offset of the line of best focus from an expected position of the line of best focus within the image, and determining a position adjustment of the surface based on the offset.

Claims42

Dependent claims

A method, comprising:

(a) positioning a first surface of a coverslip overlying a sample relative to an object plane of a microscope system, wherein the sample is on an opposite side of the coverslip from the first surface;
(b) projecting a two-dimensional pattern of light onto the first surface, wherein a focal plane of the two-dimensional pattern at a position of the surface is rotated by an angle β relative to the object plane;
(c) obtaining a two-dimensional image of the pattern of light reflected from the first surface using a detector comprising an imaging sensor oriented perpendicular to a direction of propagation of the reflected pattern of light at the sensor;
(d) analyzing the image to determine a best focus position for the pattern within the image;
and
(e) determining a position of the sample relative to the object plane based on an offset of the best focus position from an expected position within the image.

3 more elements

A system, comprising:

an illumination source configured to generate illumination light;
an optical element configured to modulate the illumination light to generate a two-dimensional pattern of light;
an apparatus configured to project the two-dimensional pattern of light onto a first surface of a coverslip overlying a sample, wherein the sample is on an opposite side of the coverslip from the first surface;
a stage configured to support the sample;
a detector configured to obtain a two-dimensional image of the pattern of light reflected from the first surface and comprising an imaging sensor oriented perpendicular to a direction of propagation of the reflected pattern of light at the sensor;
and
a controller connected to the illumination source, the projection objective, the stage, and the detector,
wherein the optical element is positioned so that a focal plane of the two-dimensional pattern is rotated by an angle β relative to a plane of the surface;
and
wherein during operation of the system, the controller is configured to:
(a) position the first surface of the coverslip relative to an object plane of the system;
(b) project the two-dimensional pattern of light onto the first surface of the coverslip using the illumination source, the optical element, and the apparatus;
(c) obtain the two-dimensional image of the pattern of light reflected from the first surface of the coverslip;
(d) analyze the image to determine a best focus position of the pattern within the image;
and
(e) determine a position of the sample relative to the object plane based on an offset of the best focus position from an expected position within the image.

13 more elements

Dependencies are inferred from the claim text.

Surface sensing in optical microscopy and automated sample scanning systems

Life of this patent

Surface sensing in optical microscopy and automated sample scanning systems

Life of this patent

Abstract

Claims42