Methods and systems for selection of detection area
Inventors
CHUNG, Yao-Kuang • TAI, Chia-En • CHANG, Liang Han
Assignees
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Publication Number
US-10436776-B2
Publication Date
2019-10-08
Expiration Date
Abstract
Provided herein are methods, systems, devices, and computer-readable storage media for selecting a detection area for a well (e.g., as part of an assay plate) comprising a plurality of encoded microcarriers. In some aspects, selecting the detection area includes obtaining one or more images of the well; calculating a center of the well according to a two-dimensional coordinate system; assigning a position, according to the two-dimensional coordinate system, of a first encoded microcarrier from the plurality; determining whether a distance between the position of the first encoded microcarrier and the center of the well according to the two-dimensional coordinate system exceeds a threshold distance; and including in the detection area one or more microcarriers whose distance from the center of the well does not exceed the threshold distance, while excluding those whose distance from the center of the well exceeds the threshold.
Core Innovation
The disclosed subject matter relates to selecting a detection area in an assay-plate well that comprises a plurality of encoded microcarriers. The system obtains data representing one or more images of a well, wherein the well contains encoded microcarriers including a first subset with a first code and a first capture agent that specifically recognizes a first analyte and a second subset with a second code different from the first code and a second capture agent that specifically recognizes a second analyte. The image data are obtained using a camera with an objective and a light source.
The workflow calculates a center of the well according to a two-dimensional coordinate system based on the obtained data, and assigns a position, according to the two-dimensional coordinate system, of a first encoded microcarrier from the plurality of encoded microcarriers based on the obtained data. The system then determines whether a distance between the position of the first encoded microcarrier and the center of the well exceeds a threshold distance.
In accordance with the determination that the distance exceeds the threshold distance, the system excludes the first encoded microcarrier from a detection area. When the determination is that the distance between the position of the first encoded microcarrier and the center of the well does not exceed the threshold distance, the system includes the first encoded microcarrier in the detection area, thereby selecting the detection area. The document further describes that microcarrier codes can be decoded and that analyte detection can be performed in association with the included microcarriers, including determining an amount of an analyte bound to a microcarrier in the detection area.
Claims Coverage
The independent claims define a system, a non-transitory computer-readable storage medium, and an electronic imaging device that implement detection-area selection by computing a well center, assigning microcarrier positions, and including or excluding microcarriers based on whether their distance from the center exceeds a threshold distance. The inventive core comprises five features.
Threshold-distance based detection area selection using a two-dimensional coordinate system
Calculating a center of the well, according to a two-dimensional coordinate system, based on the obtained data; assigning a position, according to the two-dimensional coordinate system, of a first encoded microcarrier from the plurality of encoded microcarriers based on the obtained data; determining whether a distance between the position of the first encoded microcarrier and the center of the well according to the two-dimensional coordinate system exceeds a threshold distance; excluding the first encoded microcarrier from a detection area if the distance exceeds the threshold distance; including the first encoded microcarrier in the detection area and selecting the detection area if the distance does not exceed the threshold distance.
Imaging-based identification of encoded microcarriers in an assay-plate well
Obtaining data representing one or more images of a well of an assay plate positioned on the detection stage, wherein the well comprises a plurality of encoded microcarriers, and wherein the data representing the one or more images of the well are obtained by the camera using the objective and the light source.
Analyte-specific encoded microcarriers with different codes and capture agents
The plurality of encoded microcarriers comprises a first subset of one or more encoded microcarriers, wherein each microcarrier of the first subset comprises a first code and a first capture agent that specifically recognizes a first analyte; and a second subset of one or more encoded microcarriers, wherein each microcarrier of the second subset comprises a second code and a second capture agent, wherein the second code is different from the first code.
Device and processing implementation for include/exclude detection-area selection
Providing an objective, a camera, a light source, a detection stage, and one or more processors or a processing unit configured to obtain image data of a well and to execute the center calculation, microcarrier position assignment, and threshold-distance-based determination to exclude or include a microcarrier, thereby selecting the detection area.
Non-transitory computer-readable storage medium with executable include/exclude detection-area selection
A non-transitory computer-readable storage medium comprising one or more programs for execution by one or more processors, wherein the programs include instructions that cause the device to obtain image data, calculate a center of the well according to a two-dimensional coordinate system, assign a position of a first encoded microcarrier, determine whether a distance between the microcarrier position and the well center exceeds a threshold distance, and exclude or include the microcarrier in a detection area to select the detection area.
Across the independent claims, coverage centers on imaging an assay-plate well containing encoded microcarriers with different codes and analyte-specific capture agents, computing the well center in a two-dimensional coordinate system, assigning microcarrier positions, and selecting a detection area by excluding or including microcarriers according to whether their distance from the well center exceeds a threshold distance.
Stated Advantages
Reducing fluorescence MFI variation (CV%) to <5% when selecting the detection area.
Selecting the detection area by excluding microcarriers beyond a threshold distance and including microcarriers within the threshold distance.
Documented Applications
Imaging and analysis of assay-plate wells in multiplex assays, using encoded microcarriers with different codes and capture agents and selecting a detection area based on distance from the well center.
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