Direct insertion of proteins into a living cell using an atomic force microscope with a nanoneedle |
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Authors: | Ikuo Obataya Chikashi Nakamura SungWoong Han Noriyuki Nakamura Jun Miyake |
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Institution: | (1) Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, 661-0974 Hyogo, Japan;(2) Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology (TUAT), 2-24-16 Naka-cho, Koganei, 184-8588 Tokyo, Japan |
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Abstract: | We have developed a tool for directly inserting proteins into living cells by using atomic force microscopy (AFM) and an ultrathin
needle, termed a nanoneedle. The surface of the nanoneedle was modified with His-tagged proteins using nickel chelating nitrilotriaceticacid
(NTA). The fluorescent proteins, DsRed2-His6 and EGFP-His6, could be attached to and detached from the surface of the nanoneedle. These results suggest that the Ni-NTA modified nanoneedle
can successfully be used for specific delivery of proteins. The nanoneedle modified with DsRed2-His6 was able to penetrate the surface of a living HeLa cell, as confirmed by laser scanning fluorescence microscopy and monitoring
an exerting force on the nanoneedle using AFM. Force curves using the nanoneedle indicated that the needle was able to penetrate
at displacement speeds of 0.10–10 μm/s. These results suggest that this technique can be used to directly insert proteins
into living cells and is applicable for modulation or regulation of single cell activity. |
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Keywords: | AFM living cells nanoneedle LSM protein transfer |
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