FCCS measures the coincident green and red intensity fluctuations of distinct molecules that correlate if green and red labeled particles move together through a predefined confocal volume. To perform fluorescence cross-correlation spectroscopy (FCCS), samples of interest are first labeled with fluorescent probes of different colours. The FCCS setup typically includes a confocal microscope, two laser sources, and two detectors. The confocal microscope is used to focus the laser beams and collect the fluorescence signals. The signals from the detectors are then collected and recorded over time. Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy], more text. Data analysis involves cross-correlating the signals to determine the degree of correlation between the two fluorescent probes. This information can be used to extract data on the stoichiometry and binding constants of molecular complexes, as well as the timing and location of interactions within living cells.
