Noncontact Measurement of the Local Mechanical Properties of Living Cells Using Pressure Applied via a Pipette

Sánchez, Daniel; Johnson, Nick; Li, Chao; Novák, Pavel; Rheinlaender, Johannes; Zhang, Yanjun; Anand, Uma; Anand, Praveen; Gorelik, Julia; Frolenkov, Gregory I.; Benham, Christopher D.; Lab, Max J.; Ostanin, Victor P.; Schäffer, Tilman E.; Klenerman, David; Korchev, Yuri E.

doi:10.1529/biophysj.108.129551

Cited by 113 publications

(93 citation statements)

References 48 publications

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“…The first flat slope indicates a low stiffness and is limited to the submembranous cortex of the cell. In agreement with recent force measurements applying an newly developed non-AFM method (33), there is a fluidic layer beneath the plasma membrane, which is highly dynamic in terms of thickness and viscosity. The cortical cytoskeleton of vascular endothelial cells is highly dynamic (34) and the state of polymerization of cortical actin determines the structure and mechanical properties of this layer (21,35).…”

Section: Discussionsupporting

confidence: 89%

Potassium softens vascular endothelium and increases nitric oxide release

Oberleithner

Callies

Kusche-Vihrog

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

178

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In the presence of aldosterone, plasma sodium in the high physiological range stiffens endothelial cells and reduces the release of nitric oxide. We now demonstrate effects of extracellular potassium on stiffness of individual cultured bovine aortic endothelial cells by using the tip of an atomic force microscope as a mechanical nanosensor. An acute increase of potassium in the physiological range swells and softens the endothelial cell and increases the release of nitric oxide. A high physiological sodium concentration, in the presence of aldosterone, prevents these changes. We propose that the potassium effects are caused by submembranous cortical fluidization because cortical actin depolymerization induced by cytochalasin D mimics the effect of high potassium. In contrast, a low dose of trypsin, known to activate sodium influx through epithelial sodium channels, stiffens the submembranous cell cortex. Obviously, the cortical actin cytoskeleton switches from gelation to solation depending on the ambient sodium and potassium concentrations, whereas the center of the cell is not involved. Such a mechanism would control endothelial deformability and nitric oxide release, and thus influence systemic blood pressure.aldosterone ͉ blood pressure ͉ cortical actin ͉ epithelial sodium channel ͉ stiffness

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Section: Discussionsupporting

confidence: 89%

Potassium softens vascular endothelium and increases nitric oxide release

Oberleithner

Callies

Kusche-Vihrog

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

178

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“…These approaches are widely used 31,[52][53][54]56 but require an estimate for the nanopipette inner angle, which cannot be obtained directly near the nanopipette opening using SEM, and to date has usually been estimated from the outer pipette angle obtained in SEM images. 55,[57][58][59] Evidently, this approach fails if the assumption about a constant inner angle does not hold.…”

Section: Evaluation Of Existing Methods For Nanopipette Characterizationmentioning

confidence: 99%

Characterization of Nanopipettes

et al. 2016

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Nanopipettes are widely used in electrochemical and analytical techniques as tools for sizing, sequencing, sensing, delivery and imaging. For all of these applications, the response of a nanopipette is strongly affected by its geometry and surface chemistry. As the size of nanopipettes becomes smaller, precise geometric characterization is increasingly important, especially if nanopipette probes are to be used for quantitative studies and analysis. This contribution highlights the combination of data from voltage-scanning ion conductivity experiments, transmission electron microscopy (TEM) and finite element method (FEM) simulations to fully characterize nanopipette geometry and surface charge characteristics, with an accuracy not achievable using existing approaches. Indeed, it is shown that presently used methods for nanopipette characterization can lead to highly erroneous information on nanopipettes. The new approach to characterization further facilitates high-level quantification of the behavior of nanopipettes in electrochemical systems, as demonstrated herein for a scanning ion conductance microscope (SICM) setup.

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“…However, optical tweezers might damage cells, [10][11][12] and are not appropriate for detaching adherent, spread cells from surfaces. The latter also holds for glass micropipettes, [13][14][15] the oldest instrument to manipulate single organisms.…”

Section: Force-controlled Spatial Manipulation Of Viable Mammalian Cementioning

confidence: 99%

Force-controlled spatial manipulation of viable mammalian cells and micro-organisms by means of FluidFM technology

Dörig¹,

Stiefel²,

Behr³

et al. 2010

Applied Physics Letters

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The FluidFM technology uses microchanneled atomic force microscope cantilevers that are fixed to a drilled atomic force microscope cantilevers probeholder. A continuous fluidic circuit is thereby achieved extending from an external liquid reservoir, through the probeholder and the hollow cantilever to the tip aperture. In this way, both overpressure and an underpressure can be applied to the liquid reservoir and hence to the built-in fluidic circuit. We describe in this letter how standard atomic force microscopy in combination with regulated pressure differences inside the microchanneled cantilevers can be used to displace living organisms with micrometric precision in a nondestructive way. The protocol is applicable to both eukaryotic and prokaryotic cells (e.g., mammalian cells, yeasts, and bacteria) in physiological buffer. By means of this procedure, cells can also be transferred from one glass slide to another one or onto an agar medium.

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Noncontact Measurement of the Local Mechanical Properties of Living Cells Using Pressure Applied via a Pipette

Cited by 113 publications

References 48 publications

Potassium softens vascular endothelium and increases nitric oxide release

Potassium softens vascular endothelium and increases nitric oxide release

Characterization of Nanopipettes

Force-controlled spatial manipulation of viable mammalian cells and micro-organisms by means of FluidFM technology

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