System and method for iterative detection of biological molecules
First Claim
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1. A system for iterative in situ fluorescence detection of biological molecules, comprising:
- a multi-nucleotide or antibody probe; and
a fluorophore tethered to the probe by an azide-based linker;
wherein the azide-based linker is configured to be cleaved by tris(2-carboxyethyl)phosphine (TCEP) at a plurality of sequential TCEP-specific cleavage sites within said linker thereby cleaving the fluorophore from the multi-nucleotide or antibody probe upon exposure to TCEP, andwherein the on/off ratio between a fluorescent signal produced by said fluorophore measured before treatment with TCEP and a fluorescent signal produced by said fluorophore measured after treatment with TCEP is at least about 20;
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Abstract
A system for the iterative detection of biological molecules includes a probe, and a fluorophore tethered to the probe with an azide-based linker. The linker is configured to be cleaved in the presence of tris(2-carboxyethyl)phosphine (TCEP), and the on/off ratio between a signal measured before treatment with TCEP and a signal measured after treatment with TCEP is at least about 20:1.
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9 Claims
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1. A system for iterative in situ fluorescence detection of biological molecules, comprising:
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a multi-nucleotide or antibody probe; and a fluorophore tethered to the probe by an azide-based linker; wherein the azide-based linker is configured to be cleaved by tris(2-carboxyethyl)phosphine (TCEP) at a plurality of sequential TCEP-specific cleavage sites within said linker thereby cleaving the fluorophore from the multi-nucleotide or antibody probe upon exposure to TCEP, and wherein the on/off ratio between a fluorescent signal produced by said fluorophore measured before treatment with TCEP and a fluorescent signal produced by said fluorophore measured after treatment with TCEP is at least about 20;
1. - View Dependent Claims (2, 3)
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4. A method for iterative in situ fluorescence detection a biological molecule, comprising:
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incubating a sample including a target molecule with a detection system, the detection system including a multi-nucleotide or antibody probe and a fluorophore tethered to the probe by an azide-based linker comprising a plurality of TCEP-specific cleavage sites; exciting the incubated sample at a first wavelength; detecting the emission of a second wavelength from the excited, incubated sample; and contacting tris(2-carboxyethyl)phosphine (TCEP) to the excited, incubated sample, whereby the TCEP cleaves at one or more of the plurality of sequential TCEP-specific cleavage sites, thereby cleaving the fluorophore from the probe, wherein the on/off ratio between the emission of the second wavelength detected before the contacting step and an emission of the second wavelength after the contacting step is at least about 20;
1. - View Dependent Claims (5, 6, 7, 8, 9)
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Specification