TCSPC-FLIM

FLIM

Time-correlated single photon counting - fluorescent lifetime imaging microscopy (TCSPC-FLIM) allows the highly accurate quantification of the fluorescent lifetime of a fluorescent molecule, which is informative of the fluorophores interaction status in time and space. This methodology is commonly used to quantify degrees of energy transfer (FRET) between two interacting (within intermolecular distances) single fluorescent molecules with overlapping excitation and emission spectra.

TCSPC-FLIM is available for use within ESRIC on the Leica SP5 SMD gSTED microscope (see specification) equiped with a TCSPC Module and picosecond event timer and single photon avalanche detectors (SPADs). Image credit Rebecca Saleeb, Heriot-Watt University

This microscope is available at the Heriot-Watt University site, contact Ali Dun This email address is being protected from spambots. You need JavaScript enabled to view it.

FCCS

Fluorescence cross-correlation spectroscopy is a microscopic technique derived from Fluorescence correlation spectroscopy (FCS). This method is able to determine diffusion rates and concentrations of two fluorescently-tagged molecules as they diffuse through the same pre-defined volume. When two interacting molecules are investigated the rates of diffusion differ compared to non-interacting and allow for the quantification of interaction in live samples, providing both temporal and spatial information.

FCS is performed on a Leica SP5 SMD gSTED microscope (see specification) equiped with single photon avalanche detectors (SPADs) and integrated Symphotime analysis software.

This microscope is available at the Heriot-Watt University site, contact Ali Dun This email address is being protected from spambots. You need JavaScript enabled to view it.