| CPC B01L 3/502784 (2013.01) [H04N 23/45 (2023.01); B01L 2200/16 (2013.01); B01L 2300/0654 (2013.01); B01L 2300/0877 (2013.01); H04N 23/67 (2023.01)] | 13 Claims |
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1. A system comprising:
a flow cell comprising a plurality of reaction sites a fluid inlet port and a fluid outlet port, wherein:
the plurality of reaction sites comprises a plurality of sets of reaction sites;
for each set of reaction sites from the plurality of sets of reaction sites:
that set of reaction sites is a subset of the plurality of reaction sites;
that set of reaction sites has a corresponding elevation relative to a bottom of the flow cell;
each reaction site comprised by that set of reaction sites is disposed on either a bottom surface of a corresponding well or a surface above the bottom surface of the corresponding well, and is located at that set of reaction sites' corresponding elevation relative to the bottom of the flow cell; and
no reaction site from any set of reaction sites other than that set of reaction sites is located at that set of reaction sites' corresponding elevation relative to the bottom of the flow cell;
a set of cameras; and
a processor connected to the set of cameras and programmed to:
receive a plurality of unfiltered images captured by the set of cameras, wherein the plurality of unfiltered images comprises, for each set of reaction sites from the plurality of sets of reaction sites, an image corresponding to that set of reaction sites; and
for each set of reaction sites from the plurality of sets of reaction sites, determine a derived image corresponding to that set of reaction sites, wherein the derived image corresponding to that set of reaction sites is based on using differences in focus between images from the plurality of unfiltered images to remove signals from reaction sites not comprised by that set of reaction sites while retaining signals from reaction sites comprised by that set of reaction sites;
wherein:
each set of reaction sites from the plurality of sets of reaction sites is disjoint with all other sets of reaction sites from the plurality of sets of reaction sites; and
for each image from the plurality of unfiltered images, that image depicts:
light emitted from reaction sites comprised by the set of reaction sites corresponding to that image; and
light emitted from reaction sites not comprised by the set of reaction sites corresponding to that image.
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