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Specialized Nanoneedles for Intracellular Analysis

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Nanobiosensors and Nanobioanalyses

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Abstract

Here, we introduce a novel approach to the detection of intracellular molecules by measuring direct interactions with an ultrathin probe, i.e. nanoneedle, which is mounted on an atomic force microscope (AFM). Standard AFM probes were sharpened, using a focused ion beam (FIB), to form high-aspect-ratio nanoneedles, which were then specifically functionalized and inserted into live cells. The insertion could be precisely detected using the resulted force–distance AFM curves, and no effect on cell viability was observed, even after repeated insertions. In addition, thanks to the high sensitivity of the AFM, distinct intermolecular unbinding events could be analyzed, which provided real-time information on the cytoskeleton state of the live cell. Following specific coatings and functionalization of the nanoneedles, various intracellular molecules could be detected and even inserted into live cells. The results presented here demonstrate the delivery of DNA vectors and the detection of mRNA and cytoskeletal proteins in live cells. Further advances in this technology, such as new developments in molecular functionalization options and improvements in the scale and accuracy of force measurements, will open possible new fields and applications for this diverse and powerful tool.

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Authors and Affiliations

  1. Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8562, Japan

    Ryuzo Kawamura, Yaron R. Silberberg & Chikashi Nakamura

Authors
  1. Ryuzo Kawamura
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  2. Yaron R. Silberberg
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  3. Chikashi Nakamura
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Corresponding author

Correspondence to Chikashi Nakamura .

Editor information

Editors and Affiliations

  1. School of Materials Science, Japan Advanced Institute of Science and Technology, Ishikawa, Japan

    Mun'delanji C. Vestergaard

  2. Department of Physical and Environmental Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada

    Kagan Kerman

  3. Department of Chemical and Biomolecular Engineering, The Hong Kong University of Science and Technology, Kowloon, Hong Kong SAR

    I-Ming Hsing

  4. Graduate School of Engineering, Osaka University, Osaka, Japan

    Eiichi Tamiya

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Kawamura, R., Silberberg, Y.R., Nakamura, C. (2015). Specialized Nanoneedles for Intracellular Analysis. In: Vestergaard, M., Kerman, K., Hsing, IM., Tamiya, E. (eds) Nanobiosensors and Nanobioanalyses. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55190-4_14

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