Voltage controlled nano-injection system for single-cell surgery.

Manipulation and analysis of single cells is the next frontier in understanding processes that control the function and fate of cells. Herein we describe a single-cell injection platform based on nanopipettes. The system uses scanning microscopy techniques to detect cell surfaces, and voltage pulses to deliver molecules into individual cells. As a proof of concept, we injected adherent mammalian cells with fluorescent dyes.

[1]  Kit T. Rodolfa,et al.  Two-component graded deposition of biomolecules with a double-barreled nanopipette. , 2005, Angewandte Chemie.

[2]  Alan R. Dabney,et al.  Reversible cation response with a protein-modified nanopipette. , 2011, Analytical chemistry.

[3]  T. Schäffer,et al.  Comparison of scanning ion conductance microscopy with atomic force microscopy for cell imaging. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[4]  S. Bodovitz,et al.  Single cell analysis: the new frontier in 'omics'. , 2010, Trends in biotechnology.

[5]  D. Klenerman,et al.  Nanopipette delivery of individual molecules to cellular compartments for single-molecule fluorescence tracking. , 2007, Biophysical journal.

[6]  Tomaso Zambelli,et al.  FluidFM: combining atomic force microscopy and nanofluidics in a universal liquid delivery system for single cell applications and beyond. , 2009, Nano letters.

[7]  Kit T. Rodolfa,et al.  Nanoscale pipetting for controlled chemistry in small arrayed water droplets using a double-barrel pipet. , 2006, Nano letters.

[8]  C. Parish,et al.  Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester , 2007, Nature Protocols.

[9]  U. Rant,et al.  Electrically facilitated translocations of proteins through silicon nitride nanopores: conjoint and competitive action of diffusion, electrophoresis, and electroosmosis. , 2010, Nano letters.

[10]  Chikashi Nakamura,et al.  Gene expression using an ultrathin needle enabling accurate displacement and low invasiveness. , 2005, Biochemical and biophysical research communications.

[11]  Seung-Man Yang,et al.  Nanowire-based single-cell endoscopy. , 2012, Nature nanotechnology.

[12]  N. Pourmand,et al.  Ultrasensitive mycotoxin detection by STING sensors. , 2010, Biosensors & bioelectronics.

[13]  Jacob T. Robinson,et al.  Vertical silicon nanowires as a universal platform for delivering biomolecules into living cells , 2010, Proceedings of the National Academy of Sciences.

[14]  Liming Ying,et al.  The scanned nanopipette: a new tool for high resolution bioimaging and controlled deposition of biomolecules. , 2005, Physical chemistry chemical physics : PCCP.

[15]  Ronald W Davis,et al.  Label-free biosensing with functionalized nanopipette probes , 2009, Proceedings of the National Academy of Sciences.

[16]  N. Pourmand,et al.  Dynamic Control of Nanoprecipitation in a Nanopipette , 2011, ACS nano.

[17]  N. Pourmand,et al.  Voltage-controlled metal binding on polyelectrolyte-functionalized nanopores. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[18]  Michael V. Mirkin,et al.  Electrochemical attosyringe , 2007, Proceedings of the National Academy of Sciences.

[19]  Bo Yu,et al.  Nanochannel electroporation delivers precise amounts of biomolecules into living cells. , 2011, Nature nanotechnology.

[20]  Sun Weimin,et al.  Dependence of zeta potential on polyelectrolyte moving through a solid-state nanopore , 2009 .

[21]  C. Bertozzi,et al.  A cell nanoinjector based on carbon nanotubes , 2007, Proceedings of the National Academy of Sciences.

[22]  R Pepperkok,et al.  The many ways to cross the plasma membrane , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[23]  W. Lowry,et al.  Roadblocks en route to the clinical application of induced pluripotent stem cells , 2010, Journal of Cell Science.

[24]  Jaydev P. Desai,et al.  Evaluating the Effect of Force Feedback in Cell Injection , 2007, IEEE Transactions on Automation Science and Engineering.

[25]  Alexander Meissner,et al.  Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. , 2010, Cell stem cell.

[26]  Yury Gogotsi,et al.  Multifunctional carbon-nanotube cellular endoscopes. , 2011, Nature nanotechnology.

[27]  N. Pourmand,et al.  Functionalized nanopipettes: toward label-free, single cell biosensors , 2010, Bioanalytical reviews.

[28]  Dean Ho,et al.  Nanofountain-probe-based high-resolution patterning and single-cell injection of functionalized nanodiamonds. , 2009, Small.

[29]  J. Ule,et al.  Protein–RNA interactions: new genomic technologies and perspectives , 2012, Nature Reviews Genetics.