Measurement of nanoscale three-dimensional diffusion in the interior of living cells by STED-FCS
- Lanzan\uf2, Luca
- Scipioni, Lorenzo
- Di Bona, Melody
- Bianchini, Paolo
- Bizzarri, Ranieri
- Cardarelli, Francesco
- Diaspro, Alberto
- Vicidomini, Giuseppe
DOIAbstract
The observation of molecular diffusion at different spatial scales, and in particular below the optical diffraction limit (<200 nm), can reveal details of the subcellular topology and its functional organization. Stimulated-emission depletion microscopy (STED) has been previously combined with fluorescence correlation spectroscopy (FCS) to investigate nanoscale diffusion (STED-FCS). However, stimulated-emission depletion fluorescence correlation spectroscopy has only been used successfully to reveal functional organization in two-dimensional space, such as the plasma membrane, while, an efficient implementation for measurements in three-dimensional space, such as the cellular interior, is still lacking. Here we integrate the STED-FCS method...
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