Analysis and screening of cell secretion profiles
Embodiments disclose apparatus, methods and software for performing biological screening and analysis implemented using an instrument platform capable of detecting a wide variety of cell-based secretions, expressed proteins, and other cellular components. The platform may be configured for simultaneous multiplexed detection of a plurality biological components such that a large number of discrete samples may be individually sequestered and evaluated to detect or identify constituents from the samples in a highly parallelized and scalable manner.
1. A method for spatial analysis of cells and markers on a substrate, the method comprising:
(a) acquiring an image of a plurality of cells distributed on one or more sample regions of a substrate, the substrate comprising a plurality of alignment markers associated with the one or more sample regions;
(b) detecting putative alignment markers in the image;
(c) identifying a set of alignment markers from the putative alignment markers by (i) excluding false positive alignment markers, and (ii) filling in expected locations of missing alignment markers;
(d) detecting the plurality of cells in the image;
(e) outputting a first sub-image comprising indicia of locations of the set of alignment markers; and
(f) outputting a second sub-image comprising indicia of locations of the plurality of cells.
2. The method of claim 1 , wherein the image is a bright field image.
3. The method of claim 1 , wherein the locations of the set of alignment markers are indicated in the first sub-image by a color.
4. The method of claim 1 , wherein the locations of the set of the plurality of cells are indicated in the second sub-image by a color.
5. The method of claim 1 , wherein identifying the set of alignment markers comprises size gating.
6. A method for spatial analysis of cells and markers on a substrate, the method comprising:
(a) acquiring a first image of a plurality of cells distributed on one or more sample regions of a substrate, and a second image of the plurality of cells distributed on the one or more sample regions of the substrate, wherein the substrate comprises a plurality of alignment markers associated with the one or more sample regions, and wherein the first image has a higher resolution than the second image;
(b) detecting putative alignment markers in one or both of the first image and the second image;
(c) identifying a set of alignment markers from the putative alignment markers by (i) excluding false positive alignment markers, and (ii) filling in expected location s of missing alignment markers;
(d) detecting the plurality of cells in one or both of the first image and the second image;
(e) outputting a first sub-image comprising indicia of locations of the set of alignment markers; and
(f) outputting a second sub-image comprising indicia of locations of the plurality of cells.
7. The method of claim 6 , wherein the first image and the second image are bright field images.
8. The method of claim 6 , wherein the locations of the set of alignment markers are indicated in the first sub-image by a color.
9. The method of claim 6 , wherein the locations of the set of the plurality of cells are indicated in the second sub-image by a color.
10. The method of claim 6 , wherein identifying the set of alignment markers comprises size gating.