期刊
JOURNAL OF PHYSICS D-APPLIED PHYSICS
卷 50, 期 6, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/1361-6463/aa52a1
关键词
super-resolution microscopy; actin cortex; membrane-cortex interface; plasma membrane
资金
- Wolfson Imaging Centre (Christoffer Lagerholm and Esther Garcia)
- Medical Research Council [MC_UU_12010, G0902418, MC_UU_12025]
- MRC/BBSRC/EPSRC [MR/K01577X/1]
- Wellcome Trust [104924/14/Z/14]
- Wolfson Foundation
- Engineering and Physical Sciences Research Council [1230489] Funding Source: researchfish
- Medical Research Council [G0902418, MC_UU_00008/9, MC_UU_12010/9, MR/K01577X/1] Funding Source: researchfish
- MRC [MC_UU_12010/9, MC_UU_00008/9, G0902418, MR/K01577X/1] Funding Source: UKRI
Nanoscale spacing between the plasma membrane and the underlying cortical actin cytoskeleton profoundly modulates cellular morphology, mechanics, and function. Measuring this distance has been a key challenge in cell biology. Current methods for dissecting the nanoscale spacing either limit themselves to complex survey design using fixed samples or rely on diffraction-limited fluorescence imaging whose spatial resolution is insufficient to quantify distances on the nanoscale. Using dual-color super-resolution STED (stimulated-emission-depletion) microscopy, we here overcome this challenge and accurately measure the density distribution of the cortical actin cytoskeleton and the distance between the actin cortex and the membrane in live Jurkat T-cells. We found an asymmetric cortical actin density distribution with a mean width of 230 (+105/-125) nm. The spatial distances measured between the maximum density peaks of the cortex and the membrane were bi-modally distributed with mean values of 50 +/- 15 nm and 120 +/- 40 nm, respectively. Taken together with the finite width of the cortex, our results suggest that in some regions the cortical actin is closer than 10 nm to the membrane and a maximum of 20 nm in others.
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