A Microrobotic System for Simultaneous Measurement of Turgor Pressure and Cell-Wall Elasticity of Individual Growing Plant Cells

Plant growth and morphogenesis is directed by cell division and the expansion of individual cells. How the tightly controlled process of cell expansion is regulated is poorly understood. We introduce a microrobotic platform able to separately measure the turgor pressure and cell wall elasticity of individual growing, turgid cells by combining microindentation with cell compression experiments. The system independently controls two indenters with geometries at different scales. Indentation measurements are performed automatically by deforming the cells with indenters with a spatial resolution in the nanometer range while recording force and displacement. The dual-indentation technique offers a noninvasive, high-throughput method to characterize the cytomechanics of single turgid cells by separately measuring elasticity and turgor pressure. In this way, the expansion regulation of growing cells can be investigated, as demonstrated here using Lilium longiflorum pollen tubes as an example.

[1]  B. Nelson,et al.  Quantifying growth mechanics of living, growing plant cells in situ using microbotics , 2011 .

[2]  Bradley J. Nelson,et al.  Real-time automated characterization of 3D morphology and mechanics of developing plant cells , 2015, Int. J. Robotics Res..

[3]  P B Green,et al.  Growth Physics in Nitella: a Method for Continuous in Vivo Analysis of Extensibility Based on a Micro-manometer Technique for Turgor Pressure. , 1968, Plant physiology.

[4]  Ueli Grossniklaus,et al.  The pollen tube: a soft shell with a hard core. , 2013, The Plant journal : for cell and molecular biology.

[5]  Hans R. Schultz,et al.  Growth, osmotic, adjustment, and cell-wall mechanics of expanding grape leaves during water deficits , 1993 .

[6]  Cris Kuhlemeier,et al.  Cellular Force Microscopy for in Vivo Measurements of Plant Tissue Mechanics1[W][OA] , 2012, Plant Physiology.

[7]  Chengzhi Hu,et al.  Characterization of size-dependent mechanical properties of tip-growing cells using a lab-on-chip device. , 2016, Lab on a chip.

[8]  Sina Naficy,et al.  3D/4D Printing Hydrogel Composites: A Pathway to Functional Devices , 2016 .

[9]  Arun Sampathkumar,et al.  Subcellular and supracellular mechanical stress prescribes cytoskeleton behavior in Arabidopsis cotyledon pavement cells , 2014, eLife.

[10]  Joseph A. Turner,et al.  Viscoelastic properties of cell walls of single living plant cells determined by dynamic nanoindentation , 2012, Journal of experimental botany.

[11]  P. Lintilhac,et al.  An insight into cell elasticity and load-bearing ability. Measurement and theory. , 2001, Plant physiology.

[12]  R. Benkert,et al.  The turgor pressure of growing lily pollen tubes , 1997, Protoplasma.

[13]  Monika S. Doblin,et al.  Mapping nano-scale mechanical heterogeneity of primary plant cell walls , 2016, Journal of experimental botany.

[14]  William A. Beck,et al.  Determining the Osmotic Value at Incipient Plasmolysis , 1929 .

[15]  A. D. Tomos,et al.  THE PRESSURE PROBE: A Versatile Tool in Plant Cell Physiology. , 1999, Annual review of plant physiology and plant molecular biology.

[16]  E. Bayer,et al.  Elastic Domains Regulate Growth and Organogenesis in the Plant Shoot Apical Meristem , 2012, Science.

[17]  Lan Wang,et al.  Comparison of plant cell turgor pressure measurement by pressure probe and micromanipulation , 2006, Biotechnology Letters.

[18]  Chengzhi Hu,et al.  Dual-axis Cellular Force Microscope for mechanical characterization of living plant cells , 2016, 2016 IEEE International Conference on Automation Science and Engineering (CASE).

[19]  W S Peters,et al.  Does growth correlate with turgor-induced elastic strain in stems? A re-evaluation of de Vries' classical experiments. , 2001, Plant physiology.

[20]  Emeric Bron,et al.  Pectin-Induced Changes in Cell Wall Mechanics Underlie Organ Initiation in Arabidopsis , 2011, Current Biology.

[21]  B. Nelson,et al.  Massively Parallelized Pollen Tube Guidance and Mechanical Measurements on a Lab-on-a-Chip Platform , 2016, PloS one.

[22]  John O. Outwater,et al.  Ball Tonometry: A Rapid, Nondestructive Method for Measuring Cell Turgor Pressure in Thin-Walled Plant Cells , 2000, Journal of Plant Growth Regulation.

[23]  J. Boyer,et al.  Separating growth from elastic deformation during cell enlargement , 1999, Plant physiology.

[24]  I. N. Sneddon The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile , 1965 .

[25]  Anne-Lise Routier-Kierzkowska,et al.  Measuring the mechanics of morphogenesis. , 2013, Current opinion in plant biology.

[26]  Ueli Grossniklaus,et al.  Feeling the force: how pollen tubes deal with obstacles. , 2018, The New phytologist.

[27]  Clive J Roberts,et al.  Mechanical properties of epidermal cells of whole living roots of Arabidopsis thaliana: an atomic force microscopy study. , 2012, Physical review. E, Statistical, nonlinear, and soft matter physics.

[28]  Giovanni Dietler,et al.  Atomic force microscopy stiffness tomography on living Arabidopsis thaliana cells reveals the mechanical properties of surface and deep cell-wall layers during growth. , 2012, Biophysical journal.

[29]  M. Radmacher,et al.  Bacterial turgor pressure can be measured by atomic force microscopy. , 2000, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[30]  Arezki Boudaoud,et al.  In vivo analysis of local wall stiffness at the shoot apical meristem in Arabidopsis using atomic force microscopy. , 2011, The Plant journal : for cell and molecular biology.

[31]  Anne-Lise Routier-Kierzkowska,et al.  Measuring the mechanical properties of plant cells by combining micro-indentation with osmotic treatments , 2015, Journal of experimental botany.