Patent ID: 11971354
Assignee: MOLECULAR DEVICES, LLC
Field: Measurement (Instruments)
Classification: CPC G | IPC G

Claim 0:
1. A method for multiplexed time-resolved fluorescence (TRF) detection includes:
providing one or more samples comprising a first fluorescent label bound to a first analyte, a second fluorescent label bound to a second analyte, a third fluorescent label bound to a third analyte, and a fourth fluorescent label bound to a fourth analyte, wherein
the first fluorescent label comprises a transition metal chelate and emits at a first emission wavelength having a first lifetime,
the second fluorescent label comprises a lanthanide chelate and emits at a second emission wavelength having a second lifetime,
the third fluorescent label comprises an upconverting phosphor (UCP) emitting at a third emission wavelength having a third lifetime, wherein the UCP is a lanthanide-doped or transition metal-doped inorganic compound exhibiting an anti-Stokes shift, and
the fourth fluorescent label comprises a first infrared dye emitting at a fourth emission wavelength having a fourth lifetime;

irradiating the first fluorescent label with a first excitation light at a first excitation wavelength, wherein the first fluorescent label upon irradiation with the first excitation wavelength emits a first detection signal at the first emission wavelength:
irradiating the second fluorescent label with a second excitation light at a second excitation wavelength different from the first excitation wavelength, wherein the second fluorescent label upon irradiation with the second excitation wavelength emits a second detection signal at the second emission wavelength;
irradiating the third fluorescent label with a third excitation light at a third excitation wavelength different from the first excitation wavelength and different from the second excitation wavelength, wherein the third fluorescent label upon irradiation with the third excitation wavelength emits a third detection signal at the third emission wavelength;
irradiating the fourth fluorescent label with a fourth excitation light at a fourth excitation wavelength different from the first excitation wavelength, different from the second excitation wavelength, and different from the third excitation wavelength, wherein the fourth fluorescent label upon irradiation with the fourth excitation wavelength emits a fourth detection signal at the fourth emission wavelength: and
measuring as a function of time at least one intensity of at least one of the first emission wavelength, the second emission wavelength, the third emission wavelength, and the fourth emission wavelength for a first duration, and measuring at least a different one of the first emission wavelength, the second emission wavelength, the third emission wavelength, and the fourth emission wavelength for a second duration.