Spatially encoded biological assays

Patent metadata

Field	Value
Publication number	US-12297487-B2
Application number	US-202418972052-A
Country	US
Kind code	B2
Filing date	Dec 6, 2024
Priority date	Apr 5, 2010
Publication date	May 13, 2025
Grant date	May 13, 2025

Abstract

The present invention provides assays and assay systems for use in spatially encoded biological assays. The invention provides an assay system comprising an assay capable of high levels of multiplexing where reagents are provided to a biological sample in defined spatial patterns; instrumentation capable of controlled delivery of reagents according to the spatial patterns; and a decoding scheme providing a readout that is digital in nature.

First claim

The invention claimed is:

1. A composition comprising:
(i) a substrate and a plurality of capture probes, and
(ii) a tissue section, wherein the tissue section is disposed on the substrate, and each of the plurality of capture probes comprises:
(a) a target-binding domain that comprises a nucleic acid sequence that is configured to bind to at least a portion of a nucleic acid that binds to the target- binding domain;
(b) a nucleic acid sequence that identifies a location on the substrate and is not configured to hybridize to the nucleic acid that binds to the target-binding domain, and
(c) a primer binding site.

2. The composition of claim 1 , wherein the plurality of capture probes are directly immobilized on the substrate.

3. The composition of claim 1 , wherein the plurality of capture probes are indirectly immobilized on the substrate.

4. The composition of claim 1 , wherein the plurality of capture probes are immobilized in wells on the substrate.

5. The composition of claim 1 , wherein the substrate is incorporated into a flow cell.

6. The composition of claim 1 , wherein the substrate is incorporated onto a microscope slide.

7. The composition of claim 1 , wherein the target-binding domain is configured to bind to an RNA.

8. The composition of claim 1 , wherein the target-binding domain is configured to bind to a DNA.

9. The composition of claim 1 , wherein the tissue section is a fresh frozen tissue section.

10. The composition of claim 1 , wherein the tissue section is a formalin fixed paraffin embedded tissue section.

11. The composition of claim 1 , wherein the substrate further comprises a fiducial marker.

12. The composition of claim 1 , wherein the target-binding domain is a common nucleic acid sequence among the plurality of capture probes.

13. The composition of claim 1 , further comprising a gasket.

14. The composition of claim 13 , wherein the gasket is an open gasket that surrounds the tissue section.

15. The composition of claim 1 , wherein the plurality of capture probes is attached to the substrate by a 5′ end.

16. The composition of claim 1 , further comprising reagents deposited to the tissue section.

17. A composition for spatial analysis comprising:
(i) a flow cell comprising a substrate having a plurality of wells, and
(ii) a plurality of capture probes immobilized in the wells of the substrate, wherein each of the plurality of capture probes comprises:
(a) a target-binding domain that comprises a nucleic acid sequence that is configured to bind to at least a portion of a nucleic acid that binds to the target-binding domain;
(b) a nucleic acid sequence that identifies a location of the capture probe on the substrate and is not configured to hybridize to the nucleic acid that binds to the target-binding domain, and
(c) a primer binding site.

18. The composition of claim 17 , further comprising a tissue section disposed on the substrate.

19. The composition of claim 18 , further comprising an open gasket that surrounds the tissue section.

20. The composition of claim 18 , wherein the tissue section is a fresh frozen tissue section.

21. The composition of claim 18 , wherein the tissue section is a formalin fixed paraffin embedded tissue section.

22. The composition of claim 18 , further comprising reagents deposited to the tissue section.

23. The composition of claim 17 , wherein the plurality of capture probes are directly immobilized in the wells of the substrate.

24. The composition of claim 17 , wherein the plurality of capture probes are indirectly immobilized in the wells of the substrate.

25. The composition of claim 17 , wherein the target-binding domain is configured to bind to an RNA.

26. The composition of claim 17 , wherein the target-binding domain is configured to bind to a DNA.

27. The composition of claim 17 , wherein the substrate further comprises a fiducial marker.

28. The composition of claim 17 , wherein the target-binding domain is a common nucleic acid sequence among the plurality of capture probes.

29. The composition of claim 17 , wherein the plurality of capture probes is attached to the wells of the substrate by a 5′ end.

Assignees

• Prognosys Biosciences Inc

Inventors

• Chee Mark S

Classifications

• G01N2458/40

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• C12Q2600/158

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• C12Q2600/156

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• C12Q1/6858

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