Noncontact measurement of the local mechanical properties of living cells using pressure applied via a pipette

使用移液器施加压力,非接触式测量活细胞的局部机械特性

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

Abstract

Mechanosensitivity in living biological tissue is a study area of increasing importance, but investigative tools are often inadequate. We have developed a noncontact nanoscale method to apply quantified positive and negative force at defined positions to the soft responsive surface of living cells. The method uses applied hydrostatic pressure (0.1-150 kPa) through a pipette, while the pipette-sample separation is kept constant above the cell surface using ion conductance based distance feedback. This prevents any surface contact, or contamination of the pipette, allowing repeated measurements. We show that we can probe the local mechanical properties of living cells using increasing pressure, and hence measure the nanomechanical properties of the cell membrane and the underlying cytoskeleton in a variety of cells (erythrocytes, epithelium, cardiomyocytes and neurons). Because the cell surface can first be imaged without pressure, it is possible to relate the mechanical properties to the local cell topography. This method is well suited to probe the nanomechanical properties and mechanosensitivity of living cells.

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