Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy

M. P. Clausen, H. Colin-York, Falk Schneider, Christian Eggeling, Marco Fritzsche

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Resumé

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.

OriginalsprogEngelsk
Artikelnummer064002
TidsskriftJournal of Physics D: Applied Physics
Vol/bind50
Udgave nummer6
Antal sider11
ISSN0022-3727
DOI
StatusUdgivet - 2017

Fingeraftryk

cortexes
Actins
Microscopic examination
Cells
membranes
microscopy
Membranes
Cytology
density distribution
Stimulated emission
T-cells
spacing
Cell membranes
Mechanics
Diffraction
stimulated emission
Fluorescence
Color
Imaging techniques
depletion

Citer dette

Clausen, M. P. ; Colin-York, H. ; Schneider, Falk ; Eggeling, Christian ; Fritzsche, Marco. / Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy. I: Journal of Physics D: Applied Physics. 2017 ; Bind 50, Nr. 6.
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abstract = "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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Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy. / Clausen, M. P.; Colin-York, H.; Schneider, Falk; Eggeling, Christian; Fritzsche, Marco.

I: Journal of Physics D: Applied Physics, Bind 50, Nr. 6, 064002, 2017.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy

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AU - Colin-York, H.

AU - Schneider, Falk

AU - Eggeling, Christian

AU - Fritzsche, Marco

PY - 2017

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AB - 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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KW - membrane-cortex interface

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