Studies of plasma membrane mechanics and plasma membrane–cytoskeleton interactions using optical tweezers and fluorescence imaging

Abstract We use optical tweezers in conjunction with an optical position-sensing system, which spectrally filters signals generated by a trapped fluorescent microsphere to study plasma membrane (PM) mechanics and its interactions with cytoskeleton. We dynamically measure the PM tethering force on hu...

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Published inJournal of biomechanics Vol. 40; no. 2; pp. 476 - 480
Main Authors Ermilov, Sergey A, Murdock, David R, Qian, Feng, Brownell, William E, Anvari, Bahman
Format Journal Article
LanguageEnglish
Published United States Elsevier Ltd 01.01.2007
Elsevier Limited
Subjects
Advantages
Biological forces
Biomechanical Phenomena
Cell division
Cell Line
Cell Membrane - physiology
Cytoskeleton
Cytoskeleton - physiology
Diagnostic Imaging
Experiments
Fluorescence
Humans
Laser trapping
Light
Mechanical properties
Mechanics
Optical Tweezers
Physical Medicine and Rehabilitation
Proteins
Stiffness
Tether
Viscoelasticity
Viscoelasticity
Biological forces
Stiffness
Laser trapping
Tether
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Summary:Abstract We use optical tweezers in conjunction with an optical position-sensing system, which spectrally filters signals generated by a trapped fluorescent microsphere to study plasma membrane (PM) mechanics and its interactions with cytoskeleton. We dynamically measure the PM tethering force on human embryonic kidney cells that are a standard cultured cell line. Recorded tethering force vs. PM displacement profiles, revealed the tether formation process, initiated with linear deformation of the PM, followed by a nonlinear regime and terminated with the local separation of PM. Tethering force vs. displacement profiles were used to estimate tether formation force and stiffness parameter of the PM. Integration of the force–displacement profiles yielded the work of tether formation, including linear and nonlinear components. Our results demonstrate that spectral filtering of the optically trapped fluorescent microsphere image formed on the position-sensing system overcomes the artifacts introduced by the transillumination imaging and allows accurate measures of PM mechanics before and during the initial stages of tether formation.
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ISSN:0021-9290
1873-2380
DOI:10.1016/j.jbiomech.2005.12.006