A cell nanoinjector based on carbon nanotubes

Technologies for introducing molecules into living cells are vital for probing the physical properties and biochemical interactions that govern the cell's behavior. Here, we report the development of a nanoscale cell injection system (termed the nanoinjector) that uses carbon nanotubes to deliver cargo into cells. A single multiwalled carbon nanotube attached to an atomic force microscope (AFM) tip was functionalized with cargo via a disulfide-based linker. Penetration of cell membranes with this “nanoneedle” was controlled by the AFM. The following reductive cleavage of the disulfide bonds within the cell's interior resulted in the release of cargo inside the cells, after which the nanoneedle was retracted by AFM control. The capability of the nanoinjector was demonstrated by injection of protein-coated quantum dots into live human cells. Single-particle tracking was used to characterize the diffusion dynamics of injected quantum dots in the cytosol. This technique causes no discernible membrane or cell damage, and can deliver a discrete number of molecules to the cell's interior without the requirement of a carrier solvent.

[1]  Xiaolin Nan,et al.  Observation of individual microtubule motor steps in living cells with endocytosed quantum dots. , 2005, The journal of physical chemistry. B.

[2]  Jimmy Xu,et al.  Carbon nanotube probes for single-cell experimentation and assays , 2005 .

[3]  T. D. Yuzvinsky,et al.  Length control and sharpening of atomic force microscope carbon nanotube tips assisted by an electron beam , 2005 .

[4]  H. Dai,et al.  Carbon nanotubes as multifunctional biological transporters and near-infrared agents for selective cancer cell destruction. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[5]  C. Palii,et al.  Novel fluorescence assay using calcein‐AM for the determination of human erythrocyte viability and aging , 2005, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[6]  T. Chiles,et al.  Highly efficient molecular delivery into mammalian cells using carbon nanotube spearing , 2005, Nature Methods.

[7]  C. Diaz-latoud,et al.  Cytotoxic effects induced by oxidative stress in cultured mammalian cells and protection provided by Hsp27 expression. , 2005, Methods.

[8]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[9]  M. Prato,et al.  Functionalized carbon nanotubes for plasmid DNA gene delivery. , 2004, Angewandte Chemie.

[10]  Chen Chen,et al.  Using single-particle tracking to study nuclear trafficking of viral genes. , 2004, Biophysical journal.

[11]  Wei Wang,et al.  Advances toward bioapplications of carbon nanotubes , 2004 .

[12]  Philippe Rostaing,et al.  Diffusion Dynamics of Glycine Receptors Revealed by Single-Quantum Dot Tracking , 2003, Science.

[13]  T. Waldmann,et al.  Dynamic, yet structured: The cell membrane three decades after the Singer–Nicolson model , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Joel A Swanson,et al.  Drug delivery strategy utilizing conjugation via reversible disulfide linkages: role and site of cellular reducing activities. , 2003, Advanced drug delivery reviews.

[15]  J. Gilman,et al.  Nanotechnology , 2001 .

[16]  H. Dai,et al.  Noncovalent sidewall functionalization of single-walled carbon nanotubes for protein immobilization. , 2001, Journal of the American Chemical Society.

[17]  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.

[18]  R. Ruoff,et al.  Tensile loading of ropes of single wall carbon nanotubes and their mechanical properties , 2000, Physical review letters.

[19]  R. Ruoff,et al.  Strength and breaking mechanism of multiwalled carbon nanotubes under tensile load , 2000, Science.

[20]  Elazer R. Edelman,et al.  Adv. Drug Delivery Rev. , 1997 .

[21]  K. S. Iyer,et al.  Direct spectrophotometric measurement of the rate of reduction of disulfide bonds. The reactivity of the disulfide bonds of bovine -lactalbumin. , 1973, The Journal of biological chemistry.

[22]  M. Atassi,et al.  Lack of immunochemical cross-reaction between lysozyme and alpha-lactalbumin and comparison of their conformations. , 1970, Biochimica et biophysica acta.

[23]  K. Luby-Phelps,et al.  Cytoarchitecture and physical properties of cytoplasm: volume, viscosity, diffusion, intracellular surface area. , 2000, International review of cytology.

[24]  K. Bauer,et al.  Simultaneous measurement of cell cycle and apoptotic cell death. , 1998, Methods in cell biology.