Recording intracellular (IC) bioelectrical signals is central to understanding the fundamental behaviour of cells and cell networks in, for example, neural and cardiac systems1,2,3,4. The standard tool for IC recording, the patch-clamp micropipette5 is applied widely, yet remains limited in terms of reducing the tip size, the ability to reuse the pipette5 and ion exchange with the cytoplasm6. Recent efforts have been directed towards developing new chip-based tools1,2,3,4,7,8,9,10,11,12,13, including micro-to-nanoscale metal pillars7,8,9, transistor-based kinked nanowires10,11 and nanotube devices12,13. These nanoscale tools are interesting with respect to chip-based multiplexing, but, so far, preclude targeted recording from specific cell regions and/or subcellular structures. Here we overcome this limitation in a general manner by fabricating free-standing probes in which a kinked silicon nanowire with an encoded field-effect transistor detector serves as the tip end. These probes can be manipulated in three dimensions within a standard microscope to target specific cells or cell regions, and record stable full-amplitude IC action potentials from different targeted cells without the need to clean or change the tip. Simultaneous measurements from the same cell made with free-standing nanowire and patch-clamp probes show that the same action potential amplitude and temporal properties are recorded without corrections to the raw nanowire signal. In addition, we demonstrate real-time monitoring of changes in the action potential as different ion-channel blockers are applied to cells, and multiplexed recording from cells by independent manipulation of two free-standing nanowire probes.
Parpura, V. Bionanoelectronics: getting close to the action. Nature Nanotech. 7, 143–145 (2012).
Spira, M. & Hai, A. Multi-electrode array technologies for neuroscience and cardiology. Nature Nanotech. 8, 83–94 (2013).
Duan, X., Fu, T-M., Liu, J. & Lieber, C. M. Nanoelectronics–biology frontier: from nanoscopic probes for action potential recording in live cells to three-dimensional cyborg tissues. Nano Today 8, 351–373 (2013).
Dunlop, J., Bowlby, M., Peri, R., Vasilyev, D. & Arias, R. High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology. Nature Rev. Drug Discov. 7, 358–368 (2008).
Molleman, A. Patch Clamping: An Introductory Guide to Patch Clamp Electrophysiology (Wiley, 2003).
Sakmann B. & Neher E. Single-Channel Recording (Plenum Press, 1995).
Xie, C., Lin, Z., Hanson, L., Cui, Y. & Cui, B. Intracellular recording of action potentials by nanopillar electroporation. Nature Nanotech. 7, 185–190 (2012).
Robinson, J. T. et al. Vertical nanowire electrode arrays as a scalable platform for intracellular interfacing to neuronal circuits. Nature Nanotech. 7, 180–184 (2012).
Hai, A., Shappir, J. & Spira, M. E. Long-term, multisite, parallel, in-cell recording and stimulation by an array of extracellular microelectrodes. J. Neurophysiol. 104, 559–568 (2010).
Tian, B. et al. Three-dimensional, flexible nanoscale field-effect transistors as localized bioprobes. Science 329, 830–834 (2010).
Jiang, Z., Qing, Q., Xie, P., Gao, R. X. & Lieber, C. M. Kinked p–n junction nanowire probes for high spatial resolution sensing and intracellular recording. Nano Lett. 12, 1711–1716 (2012).
Duan, X. J. et al. Intracellular recordings of action potentials by an extracellular nanoscale field-effect transistor. Nature Nanotech. 7, 174–179 (2012).
Gao, R. X. et al. Outside looking in: nanotube transistor intracellular sensors. Nano Lett. 12, 3329–3333 (2012).
Tian, B. et al. Macroporous nanowire nanoelectronic scaffolds for synthetic tissues. Nature Mater. 11, 986–994 (2012).
Tian, B., Xie, P., Kempa, T. J., Bell, D. C. & Lieber, C. M. Single-crystalline kinked semiconductor nanowire superstructures. Nature Nanotech. 4, 824–829 (2009).
Bers, D. M. Cardiac excitation–contraction coupling. Nature 415, 198–205 (2002).
Zipes, D. P. & Jalife, J. Cardiac Electrophysiology: From Cell to Bedside 2nd edn (Saunders, 2009).
Chernomordik, L. V. & Kozlov, M. M. Mechanics of membrane fusion. Nature Struct. Mol. Biol. 15, 675–683 (2008).
Nicolas, J., Mura, S., Brambilla, D., Mackiewicz, N. & Couvreur, P. Design, functionalization strategies and biomedical applications of targeted biodegradable/biocompatible polymer-based nanocarriers for drug delivery. Chem. Soc. Rev. 42, 1147–1235 (2013).
Subbiah, R., Veerapandian, M., & Yun, K. S. Nanoparticles: functionalization and multifunctional applications in biomedical sciences. Curr. Med. Chem. 17, 4559–4577 (2010).
Kotov, N. A. et al. Nanomaterials for neural interfaces. Adv. Mater. 21, 3970–4004 (2009).
Hai, A., Shappir, J. & Spira, M. E. In-cell recordings by extracellular microelectrodes. Nature Methods 7, 200–202 (2010).
Szentandrassy, N. et al. Powerful technique to test selectivity of agents acting on cardiac ion channels: the action potential voltage-clamp. Curr. Med. Chem. 18, 3737–3756 (2011).
Viatchenko-Karpinski, S. et al. Intracellular Ca2+ oscillations drive spontaneous contractions in cardiomyocytes during early development. Proc. Natl Acad. Sci. USA 96, 8259–8264 (1999).
Davie, J. T. et al. Dendritic patch-clamp recording. Nature Protocols 1, 1235–1247 (2006).
Xu, L. et al. Design and synthesis of diverse functional kinked nanowire structures for nanoelectronic bioprobes. Nano Lett. 13, 746–751 (2013).
Akbarali, H. I., Wyse, D. G. & Giles, W. R. Ionic currents in single cells from human cystic artery. Circ. Res. 70, 536–545 (1992).
Chilton, L. et al. K+ currents regulate the resting membrane potential, proliferation, and contractile responses in ventricular fibroblasts and myofibroblasts. Am. J. Physiol. Heart Circ. Physiol. 288, H2931–H2939 (2005).
Clark, B. A. & Mobbs, P. Voltage-gated currents in rabbit retinal astrocytes. Eur. J. Neurosci. 6, 1406–1414 (1994).
C.M.L. acknowledges support of this work by a National Institutes of Health Director's Pioneer Award (5DP1OD003900), National Basic Research Program of China (2013CB934103), and International Science & Technology Corporation Program of China (2013DFA50840).
The authors declare no competing financial interests.
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Qing, Q., Jiang, Z., Xu, L. et al. Free-standing kinked nanowire transistor probes for targeted intracellular recording in three dimensions. Nature Nanotech 9, 142–147 (2014). https://doi.org/10.1038/nnano.2013.273
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