Analyte detection system

An analyte detection system comprising:

• a) one or more than one particle, each particle comprising a fluorescent analyte detection dye capable of being excited by light at an excitation wavelength and capable of emitting light when excited at a maximum wavelength, and two or more than two fluorescent labels in a combination of relative amounts, wherein
• each fluorescent label is capable of being excited by light of a same excitation wavelength and capable of emitting light when excited at maximum wavelengths, distinguishable from each other, respectively, and
• the maximum wavelength of emitted light of each fluorescent analyte detection dye is different from the maximum wavelength of emitted light of each of the fluorescent labels by at least 100 nm, and
• the excitation wavelength of each analyte detection dye differs by at least 100 nm from the excitation wavelength of each of the fluorescent labels and one of the excitation wavelengths is greater than about 750 nm.
• b) means for exciting the fluorescent dye;
• c) means for exciting the first and second fluorescent labels;
• d) means for detecting the emitted lights;
• and
• e) means for correlating the detected emitted lights with a particular particle under analysis.

An assay system comprising a particle having:

• a) a fluorescent analyte detection dye capable of being excited by light at a first excitation wavelength and capable of emitting light when excited;
• b) two or more than two fluorescent labels, each fluorescent label being capable of being excited by light of a same second excitation wavelength and capable of emitting light when excited at maximum wavelengths, distinguishable from each other, respectively;
• c) a first receptor;
• and
• d) an analyte, wherein
• the analyte, first receptor, and the fluorescent analyte detection dye form a fluorescent complex on the particle, and
• the emitted light of the fluorescent analyte detection dye is different from the wavelengths of emitted lights of each of the fluorescent labels by at least 100 nm, and
• the first and second excitation wavelengths differ by at least 100 nm and one of the excitation wavelengths is greater than about 750 nm.

A method for detecting an analyte on a particle comprising:

• a) moving one or more than one particle through an examination zone, each particle having a fluorescent analyte detection dye, and two or more than two fluorescent labels;
• b) directing an exciting light of a first wavelength at each particle in the examination zone;
• c) directing an exciting light of a second wavelength at each particle in the examination zone, wherein the fluorescent analyte detection dye and the fluorescent labels each produce different emitting lights, the emitting lights each having a maximum wavelength, distinguishable from each other, respectively, wherein
• the maximum wavelength of the emitted light of the fluorescent analyte detection dye differs from the maximum wavelengths of the emitted lights of each of the fluorescent labels by at least 100 nm, and wherein the wavelengths of the first and second exciting lights differ by at least 100 nm and one of the wavelengths of exciting lights is greater than about 750 nm;
• d) detecting the emitted light of the first fluorescent analyte detection dye and the emitted light of the first and second fluorescent labels;
• and
• e) correlating the detected emitted lights with the particle under analysis.

The use of a fluorescent label in a particle for detecting an analyte comprising a particle having a fluorescent label of the formula:

• 6
• wherein:
• X and Y are each independently selected from the group consisting of O, S, NR9, and CR9R10;
• R1 and R2 are each independently selected from the group consisting of H, C1-C20 alkyl, C1-C20 haloalkyl, C1-C20 alkylene, or C1-C20 haloalkylene;
• R3 is selected from the group consisting of H, halogen, OH, OR11, SR11, NR11R12, C1-C6 alkyl, C1-C6 alkylene, C3-C6 cycloalkyl, C3-C6 cycloheteroalky, C3-C6 cycloalkylene, C3-C6 cycloheteroalkylene, phenyl, biaryl, heteroaryl, or heterobiaryl, wherein the C1-C6 alkyl, C1-C6 alkylene, C3-C6 cycloalkyl, C3-C6 cycloheteroalkyl, C3-C6 cycloalkylene, C3-C6 cycloheteroalkylene, phenyl, biaryl, heteroaryl and heterobiaryl groups may be substituted with halogen, OH, C1-C4 alkyl, or C1-C4 haloalkyl;
• R4, R5, R6, and R7 are each independently selected from the group consisting of halogen, OH, C1-C4 alkyl, or C1-C4 haloalkyl, phenyl, or heteroaryl, or other aromatic substituents known to those skilled in the art;
• R8 is selected from the group consisting of C1-C4 alkyl, or C1-C4 haloalkyl;
• R9 and R10 are each independently selected from the group consisting of hydrogen, C1-C4 alkyl, or C1-C4 haloalkyl;
• R11 and R12 are each independently selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl, phenyl, biaryl, heteroaryl, or heterobiaryl, wherein the C1-C6 alky1, C1-C6 cycloalkyl, phenyl, biaryl, heteroaryl, and heterobiaryl groups may be substituted with halogen, OH, C1-C4 alkyl, or C1-C4 haloalkyl, or when R3 represents NR11R12, R11 and R12 may be taken together to form an optionally substituted C3-C6 aliphatic or C3-C6 aromatic heterocyclic ring.