Abstract
The control of cellular function and metabolic signalling in astrocytes is complex and highly regulated. Measurement of the nanoscale localisation of protein complexes may be achieved in microscopy by the detection of Förster resonant energy transfer (FRET) between protein-conjugated fluorophores. The use of ‘ensemble’ FRET/FLIM techniques to probe protein-protein interactions in intact cells using functional FRET biosensors such as PercivalHR for metabolic imaging of ATP/ADP. Here we propose the development of a novel microscope system which will enable super-resolved functional FRET measurements within the mitochondria of astrocytes using a combination of high-speed time-correlated single photon counting and single molecule localisation microscopy.
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