Photon reducing agents and compositions for fluorescence assays
First Claim
1. A composition of matter, comprising:
- a) a membrane compartment in contact with a solid surface, wherein said membrane compartment comprises at least one photon producing agent, and b) an aqueous solution with at least one photon reducing agent, wherein said aqueous solution is in contact with an outer surface of said membrane compartment, wherein said at least one photon reducing agent is substantially impermeant to said membrane compartment, wherein said at least one photon reducing agent has an absorption spectrum that overlaps with the absorption, emission or excitation spectrum of said at least one photon producing agent, and wherein said at least one photon reducing agent is present in said aqueous solution at a concentration sufficient to reduce light emitted from said aqueous solution by at least 10% compared to the light emitted of said aqueous solution in the absence of said at least one 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 further provides a method of determining bound and free analyte in a sample using at least one photon reducing agent. The present invention also provides a method of screening test chemicals in fluorescent assays using photon reducing agents. The present invention also provides compositions and kits for practicing these methods.
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Citations
48 Claims
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1. A composition of matter, comprising:
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a) a membrane compartment in contact with a solid surface, wherein said membrane compartment comprises at least one photon producing agent, and b) an aqueous solution with at least one photon reducing agent, wherein said aqueous solution is in contact with an outer surface of said membrane compartment, wherein said at least one photon reducing agent is substantially impermeant to said membrane compartment, wherein said at least one photon reducing agent has an absorption spectrum that overlaps with the absorption, emission or excitation spectrum of said at least one photon producing agent, and wherein said at least one photon reducing agent is present in said aqueous solution at a concentration sufficient to reduce light emitted from said aqueous solution by at least 10% compared to the light emitted of said aqueous solution in the absence of said at least one 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)
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34. A composition of matter, comprising:
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a) at least one first photon producing agent internalized in a living cell in optical contact with a solid surface that can pass light of a predetermined wavelength that overlaps with the excitation spectrum of a second photon producing agent to excite said second photon producing agent, and b) at least one photon reducing agent in an aqueous solution surrounding said living cell, wherein said at least one photon reducing agent has an absorption spectra that overlaps with the absorption, emission or excitation spectra of said at least one first photon producing agent, and wherein said photon reducing agent is substantially impermeant to said membrane compartment, and wherein said at least one photon reducing agent is present in said aqueous solution at a concentration sufficient to reduce light emitted from said aqueous solution by at least 10% compared to the light emitted of said aqueous solution in the absence of said at least one photon reducing agent. - View Dependent Claims (35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48)
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Specification