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US 12,716,710 B2Granted
Systems and methods of measuring sample thickness
Assignee10x Genomics, Inc.
InventorsTobias Daniel Wheeler, Augusto Manuel Tentori
Life of this patent
Estimating the term…
Abstract
A method includes receiving an image of a plurality of fiducial markers, where the plurality of fiducial markers has at least one predetermined height relative to a substrate. The method also includes determining at least one optical metric associated with the plurality of fiducial markers within the image, and determining a calibration model based on the at least one predetermined height and the at least one optical metric.
Claims20
A method comprising:
- receiving, by one or more processors, an image of a plurality of fiducial markers on a substrate, wherein the image is acquired using an image sensor, and wherein the plurality of fiducial markers has at least one predetermined height relative to the substrate;
- processing, by the one or more processors, the image to obtain at least one optical metric associated with the plurality of fiducial markers within the image, the at least one optical metric comprising a defocused diameter of at least one of the plurality of fiducial markers when imaging at a reference focal plane;
- generating, by the one or more processors, a calibration model based on the at least one predetermined height and the at least one optical metric, the calibration model configured to provide a calibrated height or thickness of a biological sample when the calibration model is applied to one or more optical metrics obtained from an image of the biological sample with fiducial markers disposed thereon;
- and
- storing, by the one or more processors, the calibration model in a non-transitory computer-readable medium.
2 more elements
A method performed by one or more processors, the method comprising:
- accessing a calibration model from a database, the calibration model configured to provide a calibrated height or thickness when applied to an optical metric associated with a fiducial marker;
- receiving an image of a plurality of fiducial markers disposed on a biological sample, wherein the biological sample is positioned on a substrate, and wherein the image is acquired at a reference focal plane using an image sensor;
- processing the image to obtain at least one optical metric associated with the plurality of fiducial markers, the at least one optical metric comprising a defocused diameter of at least one of the plurality of fiducial markers when imaging the reference focal plane;
- and
- applying the calibration model to the at least one optical metric to obtain one or more measured heights or thicknesses of the biological sample.
2 more elements
A method comprising:
- providing a calibration slide, the calibration slide comprising a substrate having a calibration material disposed thereon, wherein the calibration material comprises at least one surface having a plurality of predetermined heights relative to the substrate;
- positioning a first plurality of fluorescent markers on the at least one surface;
- acquiring a first image of the plurality of fluorescent markers;
- determining, from the first image, at least one optical metric associated with the plurality of fluorescent markers within the first image;
- and
- determining a calibration model based on the at least one predetermined height and the at least one optical metric.
3 more elements
Dependencies are inferred from the claim text.