infocus #65 March 2022 Atomic Force Microscopy combined with Fluorescence Lifetime Imaging Microscopy (AFM+FLIM): a powerful approach to explore the structure and dynamics of biological membranes

Atomic force microscopy (AFM) and fluorescence lifetime imaging microscopy (FLIM) are two powerful techniques that are often used in the biophysical community as a means of characterising biological membranes.

DOI: 10.22443/rms.inf.1.219

On its own AFM can report on the high-resolution 3-D structure and mechanical properties of biological samples adhered to solid supports. FLIM provides information on the position and local environment of fluorescent pigments within samples.

When used in combination, these two methodologies can provide a range of opportunities for investigating biological membranes where the position and local interactions between molecules may result in changes to fluorescence, or, in the example of light-regulated processes, changes to membrane/protein functionality.

Here we present an application of combined AFM and FLIM to characterise the structure and photophysics of light-harvesting membranes, as well as using video-speed FLIM measurements to investigate membrane dynamics. This approach may be applied to a wide range of biological samples.

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