Abstract
By analyzing spatio-temporal correlations of fluorescence intensity fluctuations from raster-scanned microscopy images, raster image correlation spectroscopy (RICS) provides spatially resolved information on fast molecular dynamics such as protein diffusion or receptor-ligand interactions inside living cells and tissues [1]. Conventional RICS can only distinguish larger regions of interest, however, and requires low fluorophore concentrations in the nanomolar range, due to its diffraction-limited optical resolution. We have recently combined RICS with stimulated emission depletion (STED) microscopy to remove these limitations [2]. STED-RICS yields an enhanced multiplexing capability due to the increase in spatial resolution and can accommodate up to ∼100-fold higher fluorophore concentrations.[1] Rossow, M.J.; Sasaki, J. M.; Digman, M. A.; Gratton, E., Nat. Protoc. 2010, 5, 1761-1774.[2] Hedde, P. N.; Dorlich, R. M.; Blomley, R.; Gradl, D.; Oppong, E.; Cato, A. C.; Nienhaus, G. U., Nat Commun. 2013, 4, 2093.
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