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US 10,294,510 B2Granted
High-throughput and highly multiplexed imaging with programmable nucleic acid probes
AssigneePresident and Fellows of Harvard College
InventorsPeng Yin, Sarit Agasti, Xi Chen, Ralf Jungmann
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Abstract
The present invention provides, inter alia, methods and compositions for imaging, at high spatial resolution, targets of interest.
Claims71
A method 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 docking strand, and wherein target-specific binding partners of different specificity are linked to different docking strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) contacting the sample with labeled imager strands bind to a docking strand,
- (4) optionally removing unbound labeled imager strands,
- (5) imaging the sample to detect bound labeled imager strands,
- (6) extinguishing signal from the bound labeled imager strand, and
- (7) repeating at least some of steps (3)-(6) at least once with a labeled imager strand having a unique composition relative to at least one other labeled imager strand.
4 more elements
A method 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 docking strand, and wherein target-specific binding partners of different specificity are linked to different docking strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) contacting the sample with labeled imager strands that bind to a docking strand,
- (4) optionally removing unbound labeled imager strands,
- (5) imaging the sample to detect bound labeled imager strands,
- (6) removing the bound labeled imager strands from the docking strands by altering temperature and/or buffer condition, and
- (7) repeating at least some of steps (3)-(6) at least once with a labeled imager strand having a unique composition relative to at least one other labeled imager strand.
4 more elements
A method 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 docking strand, and wherein target-specific binding partners of different specificity are linked to different docking strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) contacting the sample with labeled imager strands that bind to a docking strand,
- (4) optionally removing unbound labeled imager strands,
- (5) imaging the sample to detect bound labeled imager strands,
- (6) inactivating the bound labeled imager strands, by removing or modifying their signal-emitting moieties without removing the imager strand in its entirety, and
- (7) repeating at least some of steps (3)-(6) at least once with a labeled imager strand having a unique composition relative to at least one other labeled imager strand.
4 more elements
A method 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 docking strand, and wherein target-specific binding partners of different specificity are linked to different docking strands,
- (2) optionally removing unbound target-specific binding partners,
- (3) contacting the sample with labeled imager strands having a nucleotide sequence
- that is complementary to a docking strand and stably binding the imager strands to the complementary docking strands,
- (4) optionally removing unbound labeled imager strands,
- (5) imaging the sample to detect location and number of bound labeled imager strands,
- (6) optionally extinguishing signal from the bound labeled imager strand, and
- (7) optionally repeating at least some of steps (3)-(6) at least once with a labeled imager strand having a unique nucleotide sequence relative to at least one other labeled imager strand.
5 more elements
Dependencies are inferred from the claim text.