The influence of lateral forces on the cell stiffness measurement by optical tweezers vertical indentation

Ndoye, Fatou; Yousafzai, Muhammad Sulaiman; Coceano, Giovanna; Bonin, Serena; Scoles, G.; Ka, Oumar; Niemela, Joseph; Cojoc, Dan

doi:10.1080/15599612.2016.1149896

Cited by 12 publications

(8 citation statements)

References 32 publications

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“…By pulling membrane tethers, OTs experiments have demonstrated that certain cell‐types exhibit lower stiffness when compared against indentation. [ 26 ] Similar experiments have demonstrated that pulling can be used to differentiate between cell types, [ 26 ] substrate‐dependent properties, [ 27 ] and drug‐dependent changes. [ 13 ] While it is hard to directly compare the viscoelastic properties of cells across different techniques, force directionality, i.e., pushing versus pulling, should also be considered.…”

Section: Resultsmentioning

confidence: 99%

An Acoustic Platform for Single‐Cell, High‐Throughput Measurements of the Viscoelastic Properties of Cells

Romanov

Silvani

Zhu

et al. 2020

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Cellular processes including adhesion, migration, and differentiation are governed by the distinct mechanical properties of each cell. Importantly, the mechanical properties of individual cells can vary depending on local physical and biochemical cues in a time‐dependent manner resulting in significant inter‐cell heterogeneity. While several different methods have been developed to interrogate the mechanical properties of single cells, throughput to capture this heterogeneity remains an issue. Here, single‐cell, high‐throughput characterization of adherent cells is demonstrated using acoustic force spectroscopy (AFS). AFS works by simultaneously, acoustically driving tens to hundreds of silica beads attached to cells away from the cell surface, allowing the user to measure the stiffness of adherent cells under multiple experimental conditions. It is shown that cells undergo marked changes in viscoelasticity as a function of temperature, by altering the temperature within the AFS microfluidic circuit between 21 and 37 °C. In addition, quantitative differences in cells exposed to different pharmacological treatments specifically targeting the membrane–cytoskeleton interface are shown. Further, the high‐throughput format of the AFS is utilized to rapidly probe, in excess of 1000 cells, three different cell lines expressing different levels of a mechanosensitive protein, Piezo1, demonstrating the ability to differentiate between cells based on protein expression levels.

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Section: Resultsmentioning

confidence: 99%

An Acoustic Platform for Single‐Cell, High‐Throughput Measurements of the Viscoelastic Properties of Cells

Romanov

Silvani

Zhu

et al. 2020

Small

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show abstract

“…Less emphasis has been placed on understanding the variation of viscoelastic properties as a function of force directionality. By pulling membrane tethers, optical tweezers experiments have demonstrated that certain cell-types exhibit lower stiffness when compared against indentation 26 . Similar experiments have demonstrated that pulling can be used to differentiate between cell types 26 , substrate-dependent properties 27 and drug-dependent changes 13 .…”

Section: Methodology For Force Calibration and Cell Viscoelasticity Mmentioning

confidence: 99%

“…By pulling membrane tethers, optical tweezers experiments have demonstrated that certain cell-types exhibit lower stiffness when compared against indentation 26 . Similar experiments have demonstrated that pulling can be used to differentiate between cell types 26 , substrate-dependent properties 27 and drug-dependent changes 13 . While it is hard to directly compare the viscoelastic properties of cells across different techniques, force-directionality, ie pushing vs pulling, should also be considered.…”

Section: Methodology For Force Calibration and Cell Viscoelasticity Mmentioning

confidence: 99%

An acoustic platform for single-cell, high-throughput measurements of the viscoelastic properties of cells

Romanov

Silvani

Zhu

et al. 2020

Preprint

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Cellular processes including adhesion, migration and differentiation are governed by the distinct mechanical properties of each cell. Importantly, the mechanical properties of individual cells can vary depending on local physical and biochemical cues in a time-dependent manner resulting in significant inter-cell heterogeneity. While several different methods have been developed to interrogate the mechanical properties of single cells, throughput to capture this heterogeneity remains an issue. While new high-throughput techniques are slowly emerging, they are primarily aimed at characterizing cells in suspension, whereas high-throughput measurements of adherent cells have proven to be more challenging. Here, we demonstrate single-cell, high-throughput characterization of adherent cells using acoustic force spectroscopy. We demonstrate that cells undergo marked changes in viscoelasticity as a function of temperature, the measurements of which are facilitated by a closed microfluidic culturing environment that can rapidly change temperature between 21 °C and 37 °C. In addition, we show quantitative differences in cells exposed to different pharmacological treatments specifically targeting the membrane-cytoskeleton interface. Further, we utilize the high-throughput format of the AFS to rapidly probe, in excess of 1000 cells, three different cell-lines expressing different levels of a mechanosensitive protein, Piezo1, demonstrating the ability to differentiate between cells based on protein expression levels.

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“…While AFM can perform this task using forces typically higher than 10 pN, the great benefit of optical manipulation is that the achievable forces complete the range of AFM reaching down to even a few tenths of a pN. Optical tweezers have been applied successfully earlier to measure cells’ Young’s modulus by trapping microbeads of various diameters and pushing them against the cells in an axial direction [ 20 , 22 , 23 , 24 ]. These cell indentation experiments use optical forces of less than 10 pN combined with a larger contact surface radius than a typical AFM tip ( r ≈ 1 μm vs. r ≈ 10 nm), which allows only small indentations, and consequently only smaller Young’s moduli can be measured.…”

Section: Introductionmentioning

confidence: 99%

Single-Cell Elasticity Measurement with an Optically Actuated Microrobot

et al. 2020

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A cell elasticity measurement method is introduced that uses polymer microtools actuated by holographic optical tweezers. The microtools were prepared with two-photon polymerization. Their shape enables the approach of the cells in any lateral direction. In the presented case, endothelial cells grown on vertical polymer walls were probed by the tools in a lateral direction. The use of specially shaped microtools prevents the target cells from photodamage that may arise during optical trapping. The position of the tools was recorded simply with video microscopy and analyzed with image processing methods. We critically compare the resulting Young’s modulus values to those in the literature obtained by other methods. The application of optical tweezers extends the force range available for cell indentations measurements down to the fN regime. Our approach demonstrates a feasible alternative to the usual vertical indentation experiments.

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The influence of lateral forces on the cell stiffness measurement by optical tweezers vertical indentation

Cited by 12 publications

References 32 publications

An Acoustic Platform for Single‐Cell, High‐Throughput Measurements of the Viscoelastic Properties of Cells

An Acoustic Platform for Single‐Cell, High‐Throughput Measurements of the Viscoelastic Properties of Cells

An acoustic platform for single-cell, high-throughput measurements of the viscoelastic properties of cells

Single-Cell Elasticity Measurement with an Optically Actuated Microrobot

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