Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy
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 themselv...
| Main Authors: | , , , , |
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| Format: | Journal article |
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Institute of Physics
2017
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| _version_ | 1826306845420027904 |
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| author | Clausen, M Colin-York, H Schneider, F Eggeling, C Fritzsche, M |
| author_facet | Clausen, M Colin-York, H Schneider, F Eggeling, C Fritzsche, M |
| author_sort | Clausen, M |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T06:54:03Z |
| format | Journal article |
| id | oxford-uuid:fd858caa-b120-44d4-97c0-87843640d04d |
| institution | University of Oxford |
| last_indexed | 2024-03-07T06:54:03Z |
| publishDate | 2017 |
| publisher | Institute of Physics |
| record_format | dspace |
| spelling | oxford-uuid:fd858caa-b120-44d4-97c0-87843640d04d2022-03-27T13:29:30ZDissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopyJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:fd858caa-b120-44d4-97c0-87843640d04dSymplectic Elements at OxfordInstitute of Physics2017Clausen, MColin-York, HSchneider, FEggeling, CFritzsche, MNanoscale 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. |
| spellingShingle | Clausen, M Colin-York, H Schneider, F Eggeling, C Fritzsche, M Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title | Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title_full | Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title_fullStr | Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title_full_unstemmed | Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title_short | Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy |
| title_sort | dissecting the actin cortex density and membrane cortex distance in living cells by super resolution microscopy |
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