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US 2025/0154559 A1Published application
Methods for Multiplex Imaging Using Labeled Nucleic Acid Imaging Agents
AssigneeUltivue, Inc.
InventorsStephanie Rae Hennek, Mael Manesse, Abdul Mohammed, Xi Chen, Michael Jules Natan
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Abstract
The present application provides certain advantageous ways of conducting multiplexed imaging.
Claims30
A method to test a sample for the presence of one or more targets comprising
- (1) contacting a sample being tested for the presence of one or more targets with one or more target-specific binding partners, wherein each target-specific binding partner is linked to a nucleic acid strand and wherein target-specific binding partners of different specificity are linked to different nucleic acid strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) contacting the sample with a nonlinear amplifier strand having complementarity to a nucleic acid strand, wherein the nucleic acid strand is either a docking strand or a primer strand,
- (4) optionally removing unbound nonlinear amplifier strands,
- (5) in either one or two steps amplifying the docking strand with rolling circle amplification and contacting the sample with labeled imager strands having complementarity to the docking strand or amplified strand,
- (6) imaging the sample to detect bound labeled imager strands,
- (7) removing the bound labeled imager strands, and
- (8) optionally repeating steps (1)-(8), or any subset thereof.
5 more elements
A method to test a sample for the presence of one or more targets comprising
- (1) contacting the sample with one or more target-specific binding partners, wherein each target-specific binding partner is linked to a nucleic acid strand, directly or indirectly, and wherein target-specific binding partners of different specificity are linked to different nucleic acid strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) wherein the nucleic acid strand is a docking strand or a primer strand and if the nucleic acid is
- (a) a docking strand, optionally increasing the number of docking strands associated with each target-specific binding partner, or
- (b) a primer strand, optionally associating more than one docking strand with the primer strand,
- (4) contacting the sample with labeled imager strands capable of binding a docking strand, directly or indirectly,
- (5) optionally removing unbound labeled imager strands,
- (6) imaging the sample to detect bound labeled imager strands, and
- (7) optionally extinguishing signal from the bound labeled imager strand;
- (8) optionally repeating steps (1)-(7) or any subset thereof.
7 more elements
A composition comprising:
- a sample bound to more than one target-specific binding partners, each binding partner bound to a nucleic acid strand and at least one docking strand stably bound to a labeled imager strand, directly or indirectly, wherein the nucleic acid strand is a docking strand or a primer strand if the nucleic acid is
- (a) a docking strand, increasing the number of docking strands associated with each target-specific binding partner, or
- (b) a primer strand, associating more than one docking strand with the primer strand.
A composition comprising
- (1) a label,
- (2) a first nucleic acid domain, a second nucleic acid domain, and a third nucleic acid domain, wherein each nucleic acid domain is from 1 to 9 nucleotides long,
- (3) a first linking moiety linking the first nucleic acid domain and the second nucleic acid domain and
- (4) a second linking moiety linking the second nucleic acid domain and the third nucleic acid domain,
- wherein both linking moieties are independently chosen from (a) an abasic site with an intact phosphodiester backbone, (b) a linker cleavable by a nucleic acid glycosylase, (c) non-natural nucleotides, or (d) restriction site or a nicking site.
2 more elements
Dependencies are inferred from the claim text.