FLIM Microscopy – Fluorescence Lifetime Imaging Microscopy (FLIM) measures the spatial distribution of fluorescence lifetimes across a sample. Unlike traditional fluorescence microscopy, which measures the intensity of emitted light, FLIM creates image contrast using the decay time of fluorescence, providing an additional layer of information. Discover how our MicroPL accessory and RMS1000 FLIM capable microscopes can support your FLIM research.
FLIM Microscopy involves exciting a sample with a pulsed laser and then measuring the time it takes for the fluorescence to decay after excitation. FLIM can be used in a variety of applications including:
Materials science: FLIM is used to measure the spatial variation in charge carrier lifetimes across semiconductor materials and investigate loss mechanisms.
Biological imaging: A sample is stained with one or more fluorophores where the fluorophore lifetime is influenced by various factors, such as the local environment, quenching effects, and interactions with other molecules.
Figure 1: FLIM Imaging of mouse intestines.
FLIM Microscopy is used for lifetimes in the picoseconds to nanoseconds time range and utilises Time-Correlated Single Photon Counting (TCSPC). Phosphorescence Lifetime Imaging (PLIM) using Multichannel Scaling Photon Counting (MCS) is a complementary technique for measuring longer microsecond and millisecond lifetime images; however, both techniques are often collectively called FLIM.
Edinburgh Instruments fluorescence spectrometers and Raman Microscopes can be extended with both FLIM and PLIM capability.