A method for multiplexed fluorescence detection, the method comprising: providing a sample comprising a first fluorescent label bound to a first analyte and a second fluorescent label bound to a second analyte; irradiating the first fluorescent label with a first excitation light at a first excitation wavelength, wherein the first fluorescent label emits a first detection signal at a first emission wavelength; then, 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 emits a second detection signal at a second emission wavelength; measuring an intensity of the first detection signal at a first measurement time, wherein the intensity of the first detection signal is correlated with the amount of the first analyte in the sample; ceasing irradiating the second fluorescent label; and after ceasing irradiating the second fluorescent label, measuring an intensity of the second detection signal at a second measurement time, wherein the intensity of the second detection signal is correlated with the amount of the second analyte in the sample; wherein the first analyte and the second analyte comprise proteins or membrane-bound proteins, and further comprising at least one of: wherein one of the first analyte and the second analyte is a reference protein, and the other of the first analyte and the second analyte is an unknown protein, and further comprising normalizing the second detection signal to the first detection signal, or normalizing the first detection signal to the second detection signal; and wherein one of the first analyte and the second analyte is an unmodified protein, and the other of the first analyte and the second analyte is a modified or phosphorylated version of the protein, and further comprising calculating a ratio of the modified or phosphorylated version of the protein to the unmodified protein based on the measured intensities of the first detection signal and the second detection signal. providing a sample comprising a first fluorescent label bound to a first analyte and a second fluorescent label bound to a second analyte; irradiating the first fluorescent label with a first excitation light at a first excitation wavelength, wherein the first fluorescent label emits a first detection signal at a first emission wavelength; then, 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 emits a second detection signal at a second emission wavelength; measuring an intensity of the first detection signal at a first measurement time, wherein the intensity of the first detection signal is correlated with the amount of the first analyte in the sample; ceasing irradiating the second fluorescent label; and after ceasing irradiating the second fluorescent label, measuring an intensity of the second detection signal at a second measurement time, wherein the intensity of the second detection signal is correlated with the amount of the second analyte in the sample; wherein the first analyte and the second analyte comprise proteins or membrane-bound proteins, and further comprising at least one of: wherein one of the first analyte and the second analyte is a reference protein, and the other of the first analyte and the second analyte is an unknown protein, and further comprising normalizing the second detection signal to the first detection signal, or normalizing the first detection signal to the second detection signal; and wherein one of the first analyte and the second analyte is an unmodified protein, and the other of the first analyte and the second analyte is a modified or phosphorylated version of the protein, and further comprising calculating a ratio of the modified or phosphorylated version of the protein to the unmodified protein based on the measured intensities of the first detection signal and the second detection signal. wherein one of the first analyte and the second analyte is a reference protein, and the other of the first analyte and the second analyte is an unknown protein, and further comprising normalizing the second detection signal to the first detection signal, or normalizing the first detection signal to the second detection signal; and wherein one of the first analyte and the second analyte is an unmodified protein, and the other of the first analyte and the second analyte is a modified or phosphorylated version of the protein, and further comprising calculating a ratio of the modified or phosphorylated version of the protein to the unmodified protein based on the measured intensities of the first detection signal and the second detection signal.