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2. Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy

Author

Actis, Paolo, Maalouf, Michelle M, Kim, Hyunsung John, Lohith, Akshar, Vilozny, Boaz, Seger, R Adam, and Pourmand, Nader

Subjects
Biochemistry and Cell Biology, Genetics, Biological Sciences, Biotechnology, Stem Cell Research - Nonembryonic - Human, Stem Cell Research, Bioengineering, Human Genome, 1.1 Normal biological development and functioning, Underpinning research, Generic health relevance, Base Sequence, Biopsy, Cells, Cultured, DNA Primers, Genomics, Humans, Microscopy, Nanotechnology, Polymerase Chain Reaction, Single-Cell Analysis, nanopipette, scanning ion conductance microscopy, single-cell biopsy, electrowetting, mitochondria, next-generation sequencing, Nanoscience & Nanotechnology
Abstract

The ability to study the molecular biology of living single cells in heterogeneous cell populations is essential for next generation analysis of cellular circuitry and function. Here, we developed a single-cell nanobiopsy platform based on scanning ion conductance microscopy (SICM) for continuous sampling of intracellular content from individual cells. The nanobiopsy platform uses electrowetting within a nanopipette to extract cellular material from living cells with minimal disruption of the cellular milieu. We demonstrate the subcellular resolution of the nanobiopsy platform by isolating small subpopulations of mitochondria from single living cells, and quantify mutant mitochondrial genomes in those single cells with high throughput sequencing technology. These findings may provide the foundation for dynamic subcellular genomic analysis.

Published
2014

4. Leukozyten

Author

Gahr, M., Seger, R. A., Lentze, Michael J., editor, Schulte, Franz J., editor, Schaub, Jürgen, editor, and Spranger, Jürgen, editor

Published
2007
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6. Diagnostik

Author

Rutishauser, M., Nicolai, T., Zach, M., Leupold, W., von der Hardt, H., Wönne, R., Riedler, J., Lindemann, H., Gruber, W., Gappa, M., Frey, U., Niggemann, B., Seidenberg, J., Ratjen, F., Nikolaizik, W. H., Seger, R. A., Lau, S., Wahn, U., Rohwedder, A., Werchau, H., Berger, C., Nadal, D., Tröger, J., Stöver, B., Kotzerke, J., Betsch, B., Poets, C. F., Nüßlein, T. G., Skopnik, H., Wenzl, T. G., Rieger, C., editor, von der Hardt, H., editor, Sennhauser, F. H., editor, Wahn, U., editor, and Zach, M., editor

Published
2004
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9. Immunologie

Author

Holländer, G. A., Wagner, N., Rajewsky, K., Habermehl, P., Schwarz, K., Wolf, H. M., Zielen, S., Friedrich, W., Nadal, D., Wahn, U., Zepp, F., Wahn, V., Seger, R. A., Lentze, Michael J., editor, Schaub, Jürgen, editor, Schulte, Franz J., editor, and Spranger, Jürgen, editor

Published
2001
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10. Diagnostik

Author

Rutishauser, M., Nicolai, T., Zach, M., Leupold, W., Wönne, R., von der Hardt, H., Riedler, J., Lindemann, H., Gruber, W., Gappa, M., Frey, U., Niggemann, B., Seidenberg, J., Ratjen, F., Nikolaizik, W. H., Seger, R. A., Lau, S., Wahn, U., Rohwedder, A., Werchau, H., Tröger, J., Stöver, B., Kotzerke, J., Poets, C. F., Nüsslein, T. G., Skopnik, H., Wenzl, T. G., Rieger, C., editor, von der Hardt, H., editor, Sennhauser, F. H., editor, Wahn, U., editor, and Zach, M., editor

Published
1999
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12. Nanopipette Apparatus for Manipulating Cells

Author

Seger, R. Adam, Actis, Paolo, Vilozny, Boaz, and Pourmand, Nader

Subjects
Chemistry And Materials (General)
Abstract

Disclosed herein are methods and systems for controlled ejection of desired material onto surfaces including in single cells using nanopipettes, as well as ejection onto and into cells. Some embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller for depositing a user defined pattern on an arbitrary substrate for the purpose of controlled cell adhesion and growth. Alternate embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller and electronic control of a voltage differential in a bore of the nanopipette electroosmotically injecting material into a cell in a high-throughput manner and with minimal damage to the cell. Yet other embodiments are directed to method and system comprising functionalized nanopipettes combined with scanning ion conductance microscopy for studying molecular interactions and detection of biomolecules inside a single living cell.

Published
2017

19. Nanopore Device for Reversible Ion and Molecule Sensing or Migration

Author

Pourmand, Nader, Vilozny, Boaz, Actis, Paolo, Seger, R. Adam, and Singaram, Bakthan

Subjects
Electronics And Electrical Engineering, Atomic And Molecular Physics
Abstract

Disclosed are methods and devices for detection of ion migration and binding, utilizing a nanopipette adapted for use in an electrochemical sensing circuit. The nanopipette may be functionalized on its interior bore with metal chelators for binding and sensing metal ions or other specific binding molecules such as boronic acid for binding and sensing glucose. Such a functionalized nanopipette is comprised in an electrical sensor that detects when the nanopipette selectively and reversibly binds ions or small molecules. Also disclosed is a nanoreactor, comprising a nanopipette, for controlling precipitation in aqueous solutions by voltage-directed ion migration, wherein ions may be directed out of the interior bore by a repulsing charge in the bore.

Published
2015

22. Metalloenzymes

Author

Pollard, J. W., Danilkovitch-Miagkova, A., Minaguchi, T., Waite, K. A., Buys, T. P. H., Lam, W. L., Müller-Hermelink, H. K., Ott, G., Robb, V. A., Henske, E. P., Lynge, E., Boyd, N., Geisler, C., Seger, R., De Wolf-Peeters, C., Sagaert, X., Sheng, S., Ribatti, D., Verstovsek, S., Akin, C., Stack, M. S., Kitada, S., Gartenhaus, R., Moretti, F., Frühwald, Michael C., Mooi, W. J., Krausz, T., Kefford, R., Peikert, T., Specks, U., Tueting, T., Pföhler, C., Blask, D. E., Stevens, R. G., Nies, A. T., Gotoh, N., Tsuchida, N., Escriba, P., Singh, V., Hickey, M., Saunders, C., Xiao, G.-H., Testa, J. R., Furtwängler, R., Scholler, N., Carbone, M., Furge, K., Woude, G. F. V., Roland, W. C., Muschel, R., Hunter, K., Welch, D. R., Vaidya, K. S., Hurst, D. R., Silveira, A. C., Zang, X. A., Bari, R., Silveira, A., Szmulewitz, R., Taylor, J., Rinker-Schaffer, C., Shim, H., Plass, C., Lindsey, J. C., Clifford, S. C., Holdenrieder, S., Steinle, A., Salih, H., Brown, D. A., Breit, S. N., Bauskin, A. R., Mousses, S., Lung, M. L., Alix-Panabieres, C., Pantel, K., Djuzenova, C. S., Dalmay, T., Ahlquist, T., Lothe, R. A., Bhat, K., Setaluri, V., Rutka, J. T., Salhia, B., Cockell, K. A., Radich, J. P., Yamagishi, S.-I., Bignami, M., Verma, M., Kumar, D., Brenner, C., Zhang, Y.-W., Jamieson, D., Chi, Y.-H., Jeang, K.-T., Roninson, I., Dragani, T., Sobolev, A. S., Powers, M. V., Workmann, P., Evans, M. F., Cooper, K., Kausch, I., Doehn, C., Janz, S., Huang, C.-L., Toland, A. E., Osinaga, E., Kaye, F., Lemos, M. C., Thakker, R. V., Teh, B. T., Ponder, B. A. J., Mulligan, L. M., Gullo, C., Klein, G., Wu, X., Araten, D. J., Loeb, L. A., Cheadle, J. P., Lipsick, J., Albihn, A., Henriksson, M., Kremens, B., Germing, U., Zangemeister-Wittke, U., Simon, H.-U., Yu, Y. P., Luo, J., and Aman, P.

Published
2020

23. Metallothionein enzymes

Author

Pollard, J. W., Danilkovitch-Miagkova, A., Minaguchi, T., Waite, K. A., Buys, T. P. H., Lam, W. L., Müller-Hermelink, H. K., Ott, G., Robb, V. A., Henske, E. P., Lynge, E., Boyd, N., Geisler, C., Seger, R., De Wolf-Peeters, C., Sagaert, X., Sheng, S., Ribatti, D., Verstovsek, S., Akin, C., Stack, M. S., Kitada, S., Gartenhaus, R., Moretti, F., Frühwald, Michael C., Mooi, W. J., Krausz, T., Kefford, R., Peikert, T., Specks, U., Tueting, T., Pföhler, C., Blask, D. E., Stevens, R. G., Nies, A. T., Gotoh, N., Tsuchida, N., Escriba, P., Singh, V., Hickey, M., Saunders, C., Xiao, G.-H., Testa, J. R., Furtwängler, R., Scholler, N., Carbone, M., Furge, K., Woude, G. F. V., Roland, W. C., Muschel, R., Hunter, K., Welch, D. R., Vaidya, K. S., Hurst, D. R., Silveira, A. C., Zang, X. A., Bari, R., Silveira, A., Szmulewitz, R., Taylor, J., Rinker-Schaffer, C., Shim, H., Plass, C., Lindsey, J. C., Clifford, S. C., Holdenrieder, S., Steinle, A., Salih, H., Brown, D. A., Breit, S. N., Bauskin, A. R., Mousses, S., Lung, M. L., Alix-Panabieres, C., Pantel, K., Djuzenova, C. S., Dalmay, T., Ahlquist, T., Lothe, R. A., Bhat, K., Setaluri, V., Rutka, J. T., Salhia, B., Cockell, K. A., Radich, J. P., Yamagishi, S.-I., Bignami, M., Verma, M., Kumar, D., Brenner, C., Zhang, Y.-W., Jamieson, D., Chi, Y.-H., Jeang, K.-T., Roninson, I., Dragani, T., Sobolev, A. S., Powers, M. V., Workmann, P., Evans, M. F., Cooper, K., Kausch, I., Doehn, C., Janz, S., Huang, C.-L., Toland, A. E., Osinaga, E., Kaye, F., Lemos, M. C., Thakker, R. V., Teh, B. T., Ponder, B. A. J., Mulligan, L. M., Gullo, C., Klein, G., Wu, X., Araten, D. J., Loeb, L. A., Cheadle, J. P., Lipsick, J., Albihn, A., Henriksson, M., Kremens, B., Germing, U., Zangemeister-Wittke, U., Simon, H.-U., Yu, Y. P., Luo, J., and Aman, P.

Published
2020

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