Photon reducing agents for fluorescence assays
First Claim
1. A system for fluorescence assays, comprising:
- a) a source of excitation light, for fluorescent excitation;
b) a detector, for measuring emission; and
c) an aqueous reagent mixture, comprising;
i) a plurality of living cells in contact with a solid surface, ii) a first reagent, comprising a photon producing agent, iii) a second reagent comprising a photon reducing agent, wherein said photon reducing agent is substantially impermeant to a plasma membrane of a living mammalian cell, wherein said photon reducing agent does not specifically bind to said membrane compartment, wherein said photon reducing agent has an absorption spectrum that overlaps with the absorption, emission or excitation spectrum of said photon producing agent, and wherein said photon reducing agent is present in said aqueous reagent mixture at an amount sufficient to reduce light emitted from said aqueous reagent mixture by at least 10% compared to the light emitted from said aqueous reagent mixture in the absence of said photon reducing agent.
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Abstract
The present invention provides a method for reducing undesirable light emission from a sample using at least one photon producing agent and at least one photon reducing agent (e.g. dye-based photon reducing agents). The present invention further provides a method for reducing undesirable light emission from a sample (e.g., a biochemical or cellular sample) with at least one photon producing agent and at least one collisional quencher. The present invention also provides a method for reducing undesirable light emission from a sample (e.g., a biochemical or cellular sample) with at least one photon producing agent and at least one quencher, such as an electronic quencher. The present invention also provides a system and method of screening test chemicals in fluorescent assays using photon reducing agents. The present invention also provides compositions, pharmaceutical compositions, and kits for practicing these methods.
62 Citations
39 Claims
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1. A system for fluorescence assays, comprising:
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a) a source of excitation light, for fluorescent excitation;
b) a detector, for measuring emission; and
c) an aqueous reagent mixture, comprising;
i) a plurality of living cells in contact with a solid surface, ii) a first reagent, comprising a photon producing agent, iii) a second reagent comprising a photon reducing agent, wherein said photon reducing agent is substantially impermeant to a plasma membrane of a living mammalian cell, wherein said photon reducing agent does not specifically bind to said membrane compartment, wherein said photon reducing agent has an absorption spectrum that overlaps with the absorption, emission or excitation spectrum of said photon producing agent, and wherein said photon reducing agent is present in said aqueous reagent mixture at an amount sufficient to reduce light emitted from said aqueous reagent mixture by at least 10% compared to the light emitted from said aqueous reagent mixture in the absence of said photon reducing agent. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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Specification