In Situ Single Cell Mechanics Characterization of Yeast Cells Using Nanoneedles Inside Environmental SEM

In this study, characterization of cellular mechanics of W303 yeast cells was conducted using nanoneedles inside an environmental SEM (ESEM). This enhanced ESEM system comprises a standard ESEM instrument as a nanoimaging tool, a cooling stage as a humidity controller for cellular biology and 7 degrees of freedom and linear actuators as nanomanipulator/effector. Four types of nanoneedles were used in our experiments, i.e., silicon (Si), titanium (Ti) coated Si, and two types of tungsten (W) nanoneedles. The Si and Ti nanoneedles were fabricated using 2 N/m spring constant cantilevers. While the W nanoneedles were fabricated using 0.09 and 2 N/m spring constant cantilevers (W0.09 and W2 nanoneedles). The Si, silicon-titanium (Si-Ti), W0.09, and W2 nanoneedles are suitable to be used for local stiffness characterization of single cells. This capability can be used in future for fast disease detection since disease cells may show different cell mechanics properties as compared to the normal cells. The Si-Ti and W2 nanoneedles can penetrate the cell without cell bursting, and this could be important in single cell surgery in future to avoid cell damage.

[1]  Chikashi Nakamura,et al.  Mechanical sensing of the penetration of various nanoneedles into a living cell using atomic force microscopy. , 2005, Biosensors & bioelectronics.

[2]  R. Coppel,et al.  The malaria-infected red blood cell: Structural and functional changes , 2001, Advances in Parasitology.

[3]  Andrew E. Pelling,et al.  Local Nanomechanical Motion of the Cell Wall of Saccharomyces cerevisiae , 2004, Science.

[4]  S. Timoshenko.  Theory of Elastic Stability , 1936 .

[5]  M. Nakajima,et al.  In-situ single cell mechanical characterization of W303 Yeast cells inside Environmental-SEM , 2007, 2007 7th IEEE Conference on Nanotechnology (IEEE NANO).

[6]  George M. Pharr,et al.  On the generality of the relationship among contact stiffness, contact area, and elastic modulus during indentation , 1992 .

[7]  Subra Suresh,et al.  Computational modeling of the forward and reverse problems in instrumented sharp indentation , 2001 .

[8]  P. Dawson,et al.  Differential growth rates of Candida utilis mother and daughter cells under phased cultivation , 1980, Journal of bacteriology.

[9]  Mitchel J. Doktycz,et al.  Intracellular integration of synthetic nanostructures with viable cells for controlled biochemical manipulation , 2003 .

[10]  M. Prato,et al.  Applications of carbon nanotubes in drug delivery. , 2005, Current opinion in chemical biology.

[11]  G C Johnston,et al.  Coordination of growth with cell division in the yeast Saccharomyces cerevisiae. , 1977, Experimental cell research.

[12]  P. Couvreur,et al.  Nanoparticles in cancer therapy and diagnosis. , 2002, Advanced drug delivery reviews.

[13]  John E. Sader,et al.  Parallel beam approximation for V‐shaped atomic force microscope cantilevers , 1995 .

[14]  Richard T. Lee,et al.  Cell mechanics and mechanotransduction: pathways, probes, and physiology. , 2004, American journal of physiology. Cell physiology.

[15]  E. Sackmann,et al.  Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. , 1999, Biophysical journal.

[16]  F. Mendels,et al.  Dynamic properties of AFM cantilevers and the calibration of their spring constants , 2006 .

[17]  J. Käs,et al.  The optical stretcher: a novel laser tool to micromanipulate cells. , 2001, Biophysical journal.

[18]  K. Möller,et al.  Monitoring dynamics of electrode reactions in Li-ion batteries by in situ ESEM , 2006 .

[19]  Chikashi Nakamura,et al.  A molecular delivery system by using AFM and nanoneedle. , 2005, Biosensors & bioelectronics.

[20]  F. Arai,et al.  In situ measurement of Young's modulus of carbon nanotubes inside a TEM through a hybrid nanorobotic manipulation system , 2006, IEEE Transactions on Nanotechnology.

[21]  David Barlam,et al.  Measurement of the mechanical properties of isolated tectorial membrane using atomic force microscopy , 2006, Proceedings of the National Academy of Sciences.

[22]  R. King,et al.  Elastic analysis of some punch problems for a layered medium , 1987 .

[23]  K E Moxham,et al.  The mechanical properties of Saccharomyces cerevisiae. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Miriam B Goodman,et al.  Molecules and Mechanisms of Mechanotransduction , 2004, The Journal of Neuroscience.

[25]  Andre Boorsma,et al.  Cell wall construction in Saccharomyces cerevisiae , 2006, Yeast.

[26]  George K Stylios,et al.  Applications of nanotechnologies in medical practice. , 2005, Injury.

[27]  L. P. Hunt,et al.  Handbook of Semiconductor Silicon Technology , 2007 .

[28]  Chwee Teck Lim,et al.  Connections between single-cell biomechanics and human disease states: gastrointestinal cancer and malaria. , 2005, Acta biomaterialia.

[29]  O. Thoumine,et al.  Microplates: a new tool for manipulation and mechanical perturbation of individual cells. , 1999, Journal of biochemical and biophysical methods.

[30]  A. Gliozzi,et al.  Mechanical properties of single living cells encapsulated in polyelectrolyte matrixes. , 2006, Journal of biotechnology.

[31]  S. N. Kundra.  Toward the emergence of nanoneurosurgery: part III-nanomedicine: targeted nanotherapy, nanosurgery and progress toward the realization of nanoneurosurgery. , 2008, Neurosurgery.

[32]  A. Donald,et al.  The use of environmental scanning electron microscopy for imaging wet and insulating materials , 2003, Nature Materials.

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

[34]  V. Labhasetwar.  Nanotechnology for drug and gene therapy: the importance of understanding molecular mechanisms of delivery. , 2005, Current opinion in biotechnology.

[35]  Saltuk B. Aksu,et al.  Calibration of atomic force microscope cantilevers using piezolevers. , 2007, The Review of scientific instruments.

[36]  H. Atkinson,et al.  In Situ Environmental Scanning Electron Microscopy (ESEM) of Semi-Solid Samples , 2006 .