Biomechanics of subcellular structures by non-invasive Brillouin microscopy

Cellular biomechanics play a pivotal role in the pathophysiology of several diseases. Unfortunately, current methods to measure biomechanical properties are invasive and mostly limited to the surface of a cell. As a result, the mechanical behaviour of subcellular structures and organelles remains poorly characterised. Here, we show three-dimensional biomechanical images of single cells obtained with non-invasive, non-destructive Brillouin microscopy with an unprecedented spatial resolution. Our results quantify the longitudinal elastic modulus of subcellular structures. In particular, we found the nucleoli to be stiffer than both the nuclear envelope (p < 0.0001) and the surrounding cytoplasm (p < 0.0001). Moreover, we demonstrate the mechanical response of cells to Latrunculin-A, a drug that reduces cell stiffness by preventing cytoskeletal assembly. Our technique can therefore generate valuable insights into cellular biomechanics and its role in pathophysiology.

[1]  Carl Paterson,et al.  Spectral broadening in Brillouin imaging , 2013 .

[2]  Itay Remer,et al.  Background-free Brillouin spectroscopy in scattering media at 780  nm via stimulated Brillouin scattering. , 2016, Optics letters.

[3]  E. Mandelkow,et al.  Microtubule structure at low resolution by x-ray diffraction. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Rob Krams,et al.  Quantification of plaque stiffness by Brillouin microscopy in experimental thin cap fibroatheroma , 2015, Journal of The Royal Society Interface.

[5]  G. Charras,et al.  The cytoplasm of living cells behaves as a poroelastic material , 2013, Nature materials.

[6]  E. Sackmann,et al.  Viscoelastic properties of erythrocyte membranes in high-frequency electric fields , 1984, Nature.

[7]  Carl Paterson,et al.  Elastic suppression in Brillouin imaging by destructive interference , 2015 .

[8]  Richard T. Lee,et al.  Lamins A and C but Not Lamin B1 Regulate Nuclear Mechanics* , 2006, Journal of Biological Chemistry.

[9]  C. Lim,et al.  Mechanics of the human red blood cell deformed by optical tweezers , 2003 .

[10]  W. Kabsch,et al.  Atomic model of the actin filament , 1990, Nature.

[11]  Adam J Engler,et al.  Preparation of Hydrogel Substrates with Tunable Mechanical Properties , 2010, Current protocols in cell biology.

[12]  Vladislav V. Yakovlev,et al.  Stimulated Brillouin Scattering Microscopic Imaging , 2015, Scientific Reports.

[13]  D. C. Lin,et al.  Mechanical properties of a reversible, DNA-crosslinked polyacrylamide hydrogel. , 2004, Journal of biomechanical engineering.

[14]  J. Lammerding,et al.  Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments. , 2015, Integrative biology : quantitative biosciences from nano to macro.

[15]  J. Randall,et al.  Brillouin scattering, density and elastic properties of the lens and cornea of the eye , 1980, Nature.

[16]  J. F. Greenleaf,et al.  Magnetic resonance elastography: Non-invasive mapping of tissue elasticity , 2001, Medical Image Anal..

[17]  S. Lindsay,et al.  A general method for determination of Brillouin linewidths by correction for instrumental effects and aperture broadening : application to high-pressure diamond anvil cell experiments , 1992 .

[18]  E. Evans,et al.  Molecular maps of red cell deformation: hidden elasticity and in situ connectivity. , 1994, Science.

[19]  C. Lim,et al.  Biomechanics approaches to studying human diseases. , 2007, Trends in biotechnology.

[20]  S. Chizhik,et al.  Atomic force microscopy probing of cell elasticity. , 2007, Micron.

[21]  S. Yun,et al.  Multistage VIPA etalons for high-extinction parallel Brillouin spectroscopy , 2011, Optics express.

[22]  K. Cunningham,et al.  The role of shear stress in the pathogenesis of atherosclerosis , 2005, Laboratory Investigation.

[23]  C. Lim,et al.  AFM indentation study of breast cancer cells. , 2008, Biochemical and biophysical research communications.

[24]  Zhaokai Meng,et al.  Brillouin spectroscopy as a new method of screening for increased CSF total protein during bacterial meningitis , 2015, Journal of biophotonics.

[25]  H. Gong,et al.  The effect of the endothelial cell cortex on atomic force microscopy measurements. , 2013, Biophysical journal.

[26]  S. Yun,et al.  Confocal Brillouin microscopy for three-dimensional mechanical imaging. , 2007, Nature photonics.

[27]  William J. Polacheck,et al.  Noncontact three-dimensional mapping of intracellular hydro-mechanical properties by Brillouin microscopy , 2015, Nature Methods.

[28]  Andrea K. Bryan,et al.  Measurement of mass, density, and volume during the cell cycle of yeast , 2009, Proceedings of the National Academy of Sciences.

[29]  J. Ophir,et al.  Elastography: A Quantitative Method for Imaging the Elasticity of Biological Tissues , 1991, Ultrasonic imaging.

[30]  S. Yun,et al.  Brillouin optical microscopy for corneal biomechanics. , 2012, Investigative ophthalmology & visual science.

[31]  Bernard Nysten,et al.  Nanoscale mapping of the elasticity of microbial cells by atomic force microscopy , 2003 .

[32]  Pilhan Kim,et al.  In vivo measurement of age-related stiffening in the crystalline lens by Brillouin optical microscopy. , 2011, Biophysical journal.

[33]  C. R. Ethier,et al.  Altered mechanobiology of Schlemm’s canal endothelial cells in glaucoma , 2014, Proceedings of the National Academy of Sciences.

[34]  U. Keyser,et al.  Real-time deformability cytometry: on-the-fly cell mechanical phenotyping , 2015, Nature Methods.

[35]  Kristie J. Koski,et al.  Non-invasive determination of the complete elastic moduli of spider silks. , 2013, Nature materials.

[36]  G. Antonacci Brillouin scattering microscopy for mechanical imaging , 2015 .

[37]  Richard T. Lee,et al.  Lamin A/C deficiency causes defective nuclear mechanics and mechanotransduction. , 2004, The Journal of clinical investigation.