High-throughput and highly multiplexed imaging with programmable nucleic acid probes
First Claim
1. 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.
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Abstract
The present invention provides, inter alia, methods and compositions for imaging, at high spatial resolution, targets of interest.
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Citations
71 Claims
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1. 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.
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38. 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.
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52. 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.
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70. 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.
Specification