Probing cell biophysical behavior based on actin cytoskeleton modeling and stretching manipulation with optical tweezers

Wang, Kaiqun; Cheng, Jin; Cheng, Shuk Han; Sun, Dong

doi:10.1063/1.4819392

Cited by 19 publications

(16 citation statements)

References 19 publications

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“…Cells were first fixed with 4% paraformaldehyde and then permeabilized with 0.5% Triton X-100 for 10 min. Cells were treated with blocking buffer (10% BSA in PBS) for 20 min and the actin cytoskeleton was stained with rhodamine-phalloidin for 10 min at room temperature [9]. Confocal images (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…One optical trap was used to hold one bead in place while one optical trap was used to manipulate the opposing bead until the bead escaped from the trap. The laser power used is 0.5, 1, 1.5, 2, 2.5 and 3 W, respectively, and details of the experiments can be referenced to the work in [9]. …”

Section: Methodsmentioning

confidence: 99%

“…Red blood cells (RBCs) from patients with sickle cell trait display lower deformability than healthy RBCs [7]. Hence, the effectiveness of a certain chemotherapy drug or medicine on cells can be evaluated by quantitatively comparing the deformation behavior of cells after treatment, and this approach brings new insights into the pathogenesis and treatment of various diseases [8, 9]. …”

Section: Introductionmentioning

confidence: 99%

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Characterization of biomechanical properties of cells through dielectrophoresis-based cell stretching and actin cytoskeleton modeling

et al. 2017

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BackgroundCytoskeleton is a highly dynamic network that helps to maintain the rigidity of a cell, and the mechanical properties of a cell are closely related to many cellular functions. This paper presents a new method to probe and characterize cell mechanical properties through dielectrophoresis (DEP)-based cell stretching manipulation and actin cytoskeleton modeling.MethodsLeukemia NB4 cells were used as cell line, and changes in their biological properties were examined after chemotherapy treatment with doxorubicin (DOX). DEP-integrated microfluidic chip was utilized as a low-cost and efficient tool to study the deformability of cells. DEP forces used in cell stretching were first evaluated through computer simulation, and the results were compared with modeling equations and with the results of optical stretching (OT) experiments. Structural parameters were then extracted by fitting the experimental data into the actin cytoskeleton model, and the underlying mechanical properties of the cells were subsequently characterized.ResultsThe DEP forces generated under different voltage inputs were calculated and the results from different approaches demonstrate good approximations to the force estimation. Both DEP and OT stretching experiments confirmed that DOX-treated NB4 cells were stiffer than the untreated cells. The structural parameters extracted from the model and the confocal images indicated significant change in actin network after DOX treatment.ConclusionThe proposed DEP method combined with actin cytoskeleton modeling is a simple engineering tool to characterize the mechanical properties of cells.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of biomechanical properties of cells through dielectrophoresis-based cell stretching and actin cytoskeleton modeling

et al. 2017

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“…These data support the importance of the actin cytoskeleton in determining cancer cell stiffness and function. 90, 91 In stem cells, cell stiffness is emerging as a “mechanical biomarker” of hMSC differentiation and adipose-derived stem cell (ASC) differentiation potential. 92, 93 Cell stiffness also constitutes part of a panel of biophysical characteristics that indicate hMSC pluripotency.…”

Section: Resultsmentioning

confidence: 99%

An inverted dielectrophoretic device for analysis of attached single cell mechanics

Urbano

Clyne

2016

Lab Chip

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Dielectrophoresis (DEP), the force induced on a polarizable body by a non-uniform electric field, has been widely used to manipulate single cells in suspension and analyze their stiffness. However, most cell types do not naturally exist in suspension but instead require attachment to the tissue extracellular matrix in vivo. Cells alter their cytoskeletal structure when they attach to a substrate, which impacts cell stiffness. It is therefore critical to be able to measure mechanical properties of cells attached to a substrate. We present a novel inverted quadrupole dielectrophoretic device capable of measuring changes in the mechanics of single cells attached to a micropatterned polyacrylamide gel. The device is positioned over a cell of defined size, a directed DEP pushing force is applied, and cell centroid displacement is dynamically measured by optical microscopy. Using this device, single endothelial cells showed greater centroid displacement in response to applied DEP pushing force following actin cytoskeleton disruption by cytochalasin D. In addition, transformed mammary epithelial cell (MCF10A-NeuT) showed greater centroid displacement in response to applied DEP pushing force compared to untransformed cells (MCF10A). DEP device measurements were confirmed by showing that the cells with greater centroid displacement also had a lower elastic modulus by atomic force microscopy. The current study demonstrates that an inverted DEP device can determine changes in single attached cell mechanics on varied substrates.

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“…Other forces experienced by the cell are caused by adhesion of the leading edge, de-adhesion at the cell body and rear, and cytoskeletal contraction (10). The precise estimation of the cell protrusion force has remained a challenging problem (11)(12)(13)(14). We recently proposed the use of robotically controlled optical tweezers to trap chemoattractant-loaded biocompatible poly(lactic-co-glycolic acid) (PLGA) microparticles, which release chemoattractant molecules into a liquid environment and induce cell polarization and migration (15).…”

Section: Introductionmentioning

confidence: 99%

A Dynamic Model of Chemoattractant-Induced Cell Migration

Yang

Gou

Wang

et al. 2015

Biophysical Journal

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Cell migration refers to a directional cell movement in response to chemoattractant stimulation. In this work, we developed a cell-migration model by mimicking in vivo migration using optically manipulated chemoattractant-loaded microsources. The model facilitates a quantitative characterization of the relationship among the protrusion force, cell motility, and chemoattractant gradient for the first time (to our knowledge). We verified the correctness of the model using migrating leukemia cancer Jurkat cells. The results show that one can achieve the ideal migrating capacity by choosing the appropriate chemoattractant gradient and concentration at the leading edge of the cell.

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Probing cell biophysical behavior based on actin cytoskeleton modeling and stretching manipulation with optical tweezers

Cited by 19 publications

References 19 publications

Characterization of biomechanical properties of cells through dielectrophoresis-based cell stretching and actin cytoskeleton modeling

Characterization of biomechanical properties of cells through dielectrophoresis-based cell stretching and actin cytoskeleton modeling

An inverted dielectrophoretic device for analysis of attached single cell mechanics

A Dynamic Model of Chemoattractant-Induced Cell Migration

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