In this section
@article{Schneider:2018:10.1101/287680,
author = {Schneider, F and Waithe, D and Galiani, S and de, la Serna JB and Sezgin, E and Eggeling, C},
doi = {10.1101/287680},
journal = {Nano Letters: a journal dedicated to nanoscience and nanotechnology},
pages = {4233--4240},
title = {Nanoscale spatio-temporal diffusion modes measured by simultaneous confocal and STED imaging},
url = {http://dx.doi.org/10.1101/287680},
volume = {18},
year = {2018}
}
TY - JOUR
AB - The diffusion dynamics in the cellular plasma membrane provide crucial insights into molecular interactions, organization, and bioactivity. Beam-scanning fluorescence correlation spectroscopy combined with super-resolution stimulated emission depletion nanoscopy (scanning STED–FCS) measures such dynamics with high spatial and temporal resolution. It reveals nanoscale diffusion characteristics by measuring the molecular diffusion in conventional confocal mode and super-resolved STED mode sequentially for each pixel along the scanned line. However, to directly link the spatial and the temporal information, a method that simultaneously measures the diffusion in confocal and STED modes is needed. Here, to overcome this problem, we establish an advanced STED–FCS measurement method, line interleaved excitation scanning STED–FCS (LIESS–FCS), that discloses the molecular diffusion modes at different spatial positions with a single measurement. It relies on fast beam-scanning along a line with alternating laser illumination that yields, for each pixel, the apparent diffusion coefficients for two different observation spot sizes (conventional confocal and super-resolved STED). We demonstrate the potential of the LIESS–FCS approach with simulations and experiments on lipid diffusion in model and live cell plasma membranes. We also apply LIESS–FCS to investigate the spatiotemporal organization of glycosylphosphatidylinositol-anchored proteins in the plasma membrane of live cells, which, interestingly, show multiple diffusion modes at different spatial positions.
AU - Schneider,F
AU - Waithe,D
AU - Galiani,S
AU - de,la Serna JB
AU - Sezgin,E
AU - Eggeling,C
DO - 10.1101/287680
EP - 4240
PY - 2018///
SN - 1530-6984
SP - 4233
TI - Nanoscale spatio-temporal diffusion modes measured by simultaneous confocal and STED imaging
T2 - Nano Letters: a journal dedicated to nanoscience and nanotechnology
UR - http://dx.doi.org/10.1101/287680
UR - https://pubs.acs.org/doi/abs/10.1021/acs.nanolett.8b01190
UR - http://hdl.handle.net/10044/1/85311
VL - 18
ER -