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US 2022/0091109 A1Published application
Probes and assays for fluorescent in-situ hybridization imaging using multiplexed fluorescent switching
AssigneeApplied Materials, Inc.
InventorsZhen Li, Marc Glazer, Yun-Ching Chang
Life of this patent
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Abstract
A readout probe for use in fluorescent in-situ hybridization imaging has a targeting portion to bind to a first hybridization sequence in an encoding probe that targets an analyte in a sample, a first fluorophore to emit light at a wavelength range, and a first cleaving region between the first fluorophore and the targeting portion.
Claims17
A readout probe for use in fluorescent in-situ hybridization imaging, the readout probe comprising:
- a targeting portion to bind to a first hybridization sequence in an encoding probe that targets an analyte in a sample;
- a first fluorophore to emit light at a wavelength range;
- and
- a first cleaving region between the first fluorophore and the targeting portion.
1 more element
A readout probe for use in fluorescent in-situ hybridization imaging, the readout probe comprising:
- a targeting portion to bind to a hybridization sequence in an encoding probe that targets an analyte in a sample;
- a fluorophore to emit light at a wavelength range;
- a quencher;
- and
- a first cleaving region between the fluorophore and the quencher.
2 more elements
An assay, comprising:
- a solution;
- a first plurality of first readout probes in the solution, the first plurality of first readout probes having a first fluorophore to emit light at a first wavelength range, a first targeting portion that binds to a first hybridization sequence in a first encoding probe, and a cleaving region between the first fluorophore and the targeting portion, the cleaving region cleavable in response to a first cleaving process;
- and
- a second plurality of second readout probes in the solution, the second plurality of second readout probes having a second fluorophore to emit light at the first wavelength range and a second targeting portion that binds to a second hybridization sequence in a second encoding probe, wherein the second fluorophore is not cleavable from the second targeting portion in response to the first cleaving process.
1 more element
An assay, comprising:
- a solution;
- a plurality of readout probes in the solution, the plurality of readout probes having a fluorophore to emit light at a wavelength range, and a first targeting portion that binds to a first hybridization sequence in an encoding probe;
- and
- a plurality of quencher probes in the solution, the plurality of quencher probes having a quencher to suppress emission of light by the fluorophore and a second targeting portion that binds to second hybridization sequence in the first encoding probe, the second hybridization sequence selected such that the quencher is positioned in sufficient proximity to a readout probe bound to the encoding probe to quench the fluorophore.
1 more element
An assay, comprising:
- a solution;
- a first plurality of first readout probes in the solution, the first plurality of first readout probes having a first fluorophore to emit light at a first wavelength range, a first targeting portion that binds to a first hybridization sequence in a first encoding probe and that has a first length such that the first readout probes are configured to be washed off the first encoding probe by a washing operation;
- and
- a second plurality of second readout probes in the solution, the second plurality of second readout probes having a second fluorophore to emit light at the first wavelength range and a second targeting portion that binds to a second hybridization sequence in a second encoding probe and that has a second length that is longer than the first length such that the second readout probes are configured to remain on the second encoding probes following the washing operation.
1 more element
Dependencies are inferred from the claim text.