Membrane nanotubes drawn by optical tweezers transmit electrical signals between mammalian cells over long distances.

Biological cells continuously change shape allowing essential functions such as cell motility, vesicle-mediated release/uptake of soluble and membrane components or nanotube-mediated cell-cell communications. Here we use single cell micromanipulation to induce functional changes of cell shape for nanobiotechnological applications. Optical tweezers are focused on the plasma membrane of living cells to pull membrane nanotubes of approximately 200 nanometre diameters and 100 micrometre lengths. Upon switching off the laser tweezer membrane nanotubes relax back to the cell surface. Single-exponential relaxation times deliver local mechanical properties of cells' plasma membrane. Nanotubes pulled beyond 100 micrometre tear off and form micrometre-sized vesicles carrying functional membrane receptors and cytoplasmic signaling components. Membrane nanotubes from one cell can be contacted to adjacent cells forming via connexins intercellular electrical connections within seconds in all directions. Our method opens broad applications for multiplexing single-cell analytics to submicrometer/subfemtoliter ranges and for creating artificial intercellular signaling networks, both not attainable by current methodologies.

[1]  Simon C Watkins,et al.  Functional connectivity between immune cells mediated by tunneling nanotubules. , 2005, Immunity.

[2]  M. Toner,et al.  Microengineering of cellular interactions. , 2000, Annual review of biomedical engineering.

[3]  Helmut Grubmüller,et al.  Molecular Anatomy of a Trafficking Organelle , 2006, Cell.

[4]  W. C. Hwang,et al.  Energy of dissociation of lipid bilayer from the membrane skeleton of red blood cells. , 1997, Biophysical journal.

[5]  W. Nelson,et al.  Synapses: sites of cell recognition, adhesion, and functional specification. , 2007, Annual review of biochemistry.

[6]  W. Brownell,et al.  Membrane tether formation from outer hair cells with optical tweezers. , 2002, Biophysical journal.

[7]  S. Bromley,et al.  The immunological synapse. , 2001, Annual review of immunology.

[8]  F. Marga,et al.  Eukaryotic membrane tethers revisited using magnetic tweezers , 2007, Physical biology.

[9]  Kai Simons,et al.  Lipid Rafts As a Membrane-Organizing Principle , 2010, Science.

[10]  T. Smart,et al.  HEK293 cell line: a vehicle for the expression of recombinant proteins. , 2005, Journal of pharmacological and toxicological methods.

[11]  M. Bennett,et al.  PHYSIOLOGY OF ELECTROTONIC JUNCTIONS * , 1966, Annals of the New York Academy of Sciences.

[12]  R. Bruzzone,et al.  Connections with connexins: the molecular basis of direct intercellular signaling. , 1996, European journal of biochemistry.

[13]  E. Evans,et al.  Hidden dynamics in rapid changes of bilayer shape , 1994 .

[14]  Ofer Feinerman,et al.  Reliable neuronal logic devices from patterned hippocampal cultures , 2008 .

[15]  Horst Vogel,et al.  An integrated self-assembled nanofluidic system for controlled biological chemistries. , 2008, Angewandte Chemie.

[16]  Horst Vogel,et al.  Investigating cellular signaling reactions in single attoliter vesicles. , 2005, Journal of the American Chemical Society.

[17]  H. Gerdes,et al.  Tunneling nanotubes: A new route for the exchange of components between animal cells , 2007, FEBS letters.

[18]  Christina E. Rommel,et al.  Elasticity mapping of pore-suspending native cell membranes. , 2009, Small.

[19]  D. Davis,et al.  Membrane nanotubes: dynamic long-distance connections between animal cells , 2008, Nature Reviews Molecular Cell Biology.

[20]  Tony Pawson,et al.  Protein modules and signalling networks , 1995, Nature.

[21]  M. Bennett,et al.  Dynamics of electrical transmission at club endings on the Mauthner cells , 2004, Brain Research Reviews.

[22]  E. Evans,et al.  Giant vesicle bilayers composed of mixtures of lipids, cholesterol and polypeptides. Thermomechanical and (mutual) adherence properties. , 1986, Faraday discussions of the Chemical Society.

[23]  E. Evans,et al.  Structure and deformation properties of red blood cells: concepts and quantitative methods. , 1989, Methods in enzymology.

[24]  D A Stenger,et al.  Central neuronal synapse formation on micropatterned surfaces. , 1998, Brain research. Developmental brain research.

[25]  A. Offenhäusser,et al.  Ultrathin Coatings with Change in Reactivity over Time Enable Functional In Vitro Networks Of Insect Neurons , 2008, Advanced materials.

[26]  Hans-Hermann Gerdes,et al.  Nanotubular Highways for Intercellular Organelle Transport , 2004, Science.

[27]  K. L. Martinez,et al.  Monitoring the diffusion of single heterotrimeric G proteins in supported cell-membrane sheets reveals their partitioning into microdomains. , 2006, Journal of molecular biology.

[28]  J. Shao,et al.  Deformation and flow of membrane into tethers extracted from neuronal growth cones. , 1996, Biophysical journal.

[29]  D Kleinfeld,et al.  Controlled outgrowth of dissociated neurons on patterned substrates , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[30]  A. Ashkin,et al.  Internal cell manipulation using infrared laser traps. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[31]  O. Orwar,et al.  Chapter 15 - Complex nanotube-liposome networks. , 2009, Methods in enzymology.

[32]  J. Dai,et al.  Membrane tether formation from blebbing cells. , 1999, Biophysical journal.

[33]  Nathan M Sherer,et al.  Cytonemes and tunneling nanotubules in cell-cell communication and viral pathogenesis. , 2008, Trends in cell biology.

[34]  S. Boxer,et al.  Advances in imaging secondary ion mass spectrometry for biological samples. , 2009, Annual review of biophysics.

[35]  L. Roux,et al.  Over Astroglial Networks: a Step Further in Neuroglial and Gliovascular Interactions , 2022 .

[36]  G. Turcatti,et al.  Probing the binding domain of the NK2 receptor with fluorescent ligands: evidence that heptapeptide agonists and antagonists bind differently. , 1995, Biochemistry.

[37]  D. Davis,et al.  Membrane nanotubes facilitate long-distance interactions between natural killer cells and target cells , 2010, Proceedings of the National Academy of Sciences.

[38]  R. Hochmuth.  Solid and liquid behavior of red cell membrane. , 1982, Annual review of biophysics and bioengineering.

[39]  Nicholas G Hatsopoulos,et al.  The science of neural interface systems. , 2009, Annual review of neuroscience.

[40]  Rona Chandrawati,et al.  Polymer hydrogel capsules: en route toward synthetic cellular systems. , 2009, Nanoscale.

[41]  J. Dai,et al.  Mechanical properties of neuronal growth cone membranes studied by tether formation with laser optical tweezers. , 1995, Biophysical journal.

[42]  V. Lučić,et al.  Structural studies by electron tomography: from cells to molecules. , 2005, Annual review of biochemistry.

[43]  P. Mattila,et al.  Filopodia: molecular architecture and cellular functions , 2008, Nature Reviews Molecular Cell Biology.