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Simultaneous measurement of intracellular calcium and electrical activity from patterned neural networks in culture
Abstract
Multisite extracellular electrical activity and intracellular calcium were recorded simultaneously. Electrical signals were measured using microelectrode array substrates. A novel cell positioning technique was combined with a method for controlling neurite outgrowth, which allowed cell-electrode contacts to be established easily, thus facilitating the electrical recording. Intracellular calcium was measured optically using the indicator fluo 3. Under low-magnesium conditions, cultured rat cortical neurons showed periodic transients of fluo-3 fluorescence, which were synchronized with the periodic bursting observed electrically. The intervals between bursts could be determined by electrical stimulation through the substrate electrodes. The results suggest that functional synaptic connections are formed in the culture system.
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“Having surface topologies on the substrate, axonal outgrowth of embryonic nerve cells from rats was observed and electrical activity was recorded in a microstructure with 16 electrodes that were connected with grooves (Jimbo and Kawana, 1992; Jimbo et al., 1993)”
From A critical review of interfaces with the peripheral nervous system for the control of neuroprostheses and hybrid bionic systems (2005) · cites this paper as Jimbo et al., 1993
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“Fast spiking, caused by rapid calcium influx and efflux due to synaptic stimuli and AP propagation can been used to monitor electrophysiological activity (Jimbo et al., 1993) and connectivity in cultured neurons (Grienberger and Konnerth, 2012).”
Development of 3-D Multi-Electrode Arrays for Use with Acute Tissue Slices
Heuschkel, M. O., Wirth, C., Steidl, E.-M., & Buisson, B. (2006) · Springer eBooks · doi:10.1007/0-387-25858-2_4 · Cited by 12
“Local Field Potentials (LFP) as well as single-unit action potentials have been recorded successfully in dissociated spinal cord preparations (Gross et al., 1993; Streit et al., 2001), cortical cell cultures ( Jimbo et al., 1993; Maeda et al., 1995; Gross et al., 1997), acute retina preparations (Litke and Meister, 19…”
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Segmentation of the response of a neuronal network into clusters with similar activity.
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Self-citationMulti-Electrode Arrays: Enhancing Traditional Methods and Enabling Network Physiology
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“Early work in this field was done by Jimbo et al. (1993) .”
BackgroundProperties of the Evoked Spatio-Temporal Electrical Activity in Neuronal Assemblies
Pinato, G., Parodi, P., Bisso, A., Macrí, D., Kawana, A., Jimbo, Y., & Torre, V. (1999) · Reviews in the Neurosciences · doi:10.1515/revneuro.1999.10.3-4.279 · Cited by 10
Self-citationSahni, G., Yuan, J., & Toh, Y. (2016) · Journal of Visualized Experiments · doi:10.3791/54097 · Cited by 8
A microstructure for interfacing with neurons: the neurochip
Maher, M. P., Wright, J. A., Pine, J., & Tai, Y.-C. (1998) · Proceedings of the 20th Annual International Conference of the IEEE Engineering in Medicine and Biology Society. Vol.20 Biomedical Engineering Towards the Year 2000 and Beyond (Cat. No.98CH36286) · doi:10.1109/iembs.1998.746911 · Cited by 9
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Hattori, S., Suzurikawa, J., Kanzaki, R., Jimbo, Y., Hamaguchi, T., Takahashi, H., & Nakao, M. (2010) · doi:10.1541/ieejeiss.128.1036 · Cited by 8
Self-citationA Micro-tool for Mechanical Manipulation of in vitro Cell Arrays
Baldi, A., Fass, J., Silva, M. N. D., Odde, D. J., & Ziaie, B. (2003) · Biomedical Microdevices · doi:10.1023/a:1027353612618 · Cited by 10
Cultures of neurons on micro-electrode array in hybrid retinal implant
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“At present, this technique is being developed in several laboratories [3, 4], including ours [5], and shows great promise to become a new powerful tool for in vitro testing of experimental conditions somehow related to in vivo ones .”
Impact of microchannel width on axons for brain-on-chip applications
Vulić, K., Amos, G., Ruff, T., Kasm, R., Ihle, S. J., Küchler, J., Vörös, J., & Weaver, S. M. (2024) · bioRxiv · doi:10.1101/2024.05.16.594497 · Cited by 7
“Notably, patterning methodologies can be combined with microelectrode arrays (MEAs)[29, 30, 27], allowing for the recording and modulation of extracellular neural activity, thus providing a functional aspect to create engineered neural systems.”
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“They have also been used to study the behavior of small neuronal networks (Muller et al. 1997; Jimbo et al. 1993; Ternaux and Portalier 1993; Kuroda et al. 1995; Gopal and Gross 1996a, 1996b; Gross et al. 1997; Harsch et al. 1997; Muller et al. 1997; Misgeld et al. 1998).”
MethodThe neuro-electronic interface: measurements and model predictions
Grattarola, M., Bove, M., Verreschi, G., Martinoia, S., & Tedesco, M. (1996) · Journal of Materials Science Materials in Medicine · doi:10.1007/bf00154550 · Cited by 6
Lassers, S. B., Vakilna, Y., Tang, W. C., & Brewer, G. (2023) · Frontiers in Neural Circuits · doi:10.3389/fncir.2023.1272925 · Cited by 5
“Bursts of spikes are needed to raise the somal calcium (Jimbo et al., 1993) that is readily recorded in localized regions in vivo .”
BackgroundTonomura, W., Moriguchi, H., Jimbo, Y., & Konishi, S. (2010) · Biomedical microdevices · doi:10.1007/s10544-010-9427-0 · Cited by 5
Self-citationStochastic Simulation Model for Patterned Neural Multi-Electrode Arrays
Kahng, D.-S., Nam, Y., & Lee, D. (2007) · 2007 IEEE 7th International Symposium on BioInformatics and BioEngineering · doi:10.1109/bibe.2007.4375642 · Cited by 5
“The properties of the in vitro neural networks would provide important clues for the study of real brains as well as new opportunities of engineered intelligence [1-4].”
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Suzurikawa, J., Takahashi, H., Takayama, Y., Warisawa, S., Mitsuishi, M., Nakao, M., & Jimbo, Y. (2006) · Annual International Conference of the IEEE Engineering in Medicine and Biology Society · doi:10.1109/iembs.2006.259828 · Cited by 5
“The fluo-4 calcium imaging with cooled-CCD camera (Hamamatsu Photonics, C8800-21C) [ 11 ] was used to monitor the effect of the electrical stimulation.”
MethodSelf-citationNeurointerface-interfaces of neuronal networks to electrical circuit
Kawana, A., & Jimbo, Y. (1999) · Technical Digest. IEEE International MEMS 99 Conference. Twelfth IEEE International Conference on Micro Electro Mechanical Systems (Cat. No.99CH36291) · doi:10.1109/memsys.1999.746744 · Cited by 5
Self-citationNeuroengineering in biological and biosynthetic systems.
Buettner, H. (1995) · Current Opinion in Biotechnology · doi:10.1016/0958-1669(95)80037-9 · Cited by 5
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Perfopmance Of A Thin Film Microelectrode Array For Monitoring Electrogenic Cells In Vitro
Mohr, A., Finger, W., Fohr, K. J., Nisch, W., & Göpel, W. (1995) · International Conference on Solid-State Sensors, Actuators and Microsystems · doi:10.1109/sensor.1995.721852 · Cited by 4
Relationship Between Evoked and Spontaneous Activity in Cultured Neuronal Circuits
Kiyohara, A., Taguchi, T., & Kudoh, S. N. (2009) · IEEJ Transactions on Electronics Information and Systems · doi:10.1541/ieejeiss.129.1815 · Cited by 6
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Dynamics of networks of biological neurons: Simulation and experimental tools
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“The Living neurons reorganized complex networks on a culture dish with multi-electrodes and frequent spontaneous electrical activity was observed [1-3].”
BackgroundThe effects of the current stimulation on electrical activity in dissociated neurons
Tokuda, M., Kiyohara, A., Taguch, T., & Kudoh, S. N. (2009) · Micro-NanoMechatronics and Human Science · doi:10.1109/mhs.2009.5352068 · Cited by 3
“Complicating the temporal pattern of the spontaneous activity was reported to depend on culture days [3,5].”
BackgroundMethodCurrent Issues and Advances in Dissociated Cell Culturing on Nano-and Microfabricated Substrates
Craighead, H. G., James, C. D., & Turner, A. M. P. (2003) · Elsevier eBooks · doi:10.1016/b978-012507060-7/50026-x · Cited by 3
A microtool for in vitro cell array manipulation
Baldi, A., Fass, J., Silva, M. N. D., Odde, D. J., & Ziaie, B. (2003) · International IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology (MMB) · doi:10.1109/mmb.2002.1002309 · Cited by 2
Nam, Y., Wheeler, B. C., & Brewer, G. (2002) · 2nd Annual International IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology. Proceedings (Cat. No.02EX578) · doi:10.1109/mmb.2002.1002279 · Cited by 3
Recording of patterned neural networks
Chang, J. C., Wheeler, B. C., & Brewer, G. (1999) · Proceedings of the First Joint BMES/EMBS Conference. 1999 IEEE Engineering in Medicine and Biology 21st Annual Conference and the 1999 Annual Fall Meeting of the Biomedical Engineering Society (Cat. N · doi:10.1109/iembs.1999.802439 · Cited by 3
Thielecke, H., Stieglitz, T., Beutel, H., Matthies, T., & Meyer, J. (1998) · Proceedings of the 20th Annual International Conference of the IEEE Engineering in Medicine and Biology Society. Vol.20 Biomedical Engineering Towards the Year 2000 and Beyond (Cat. No.98CH36286) · doi:10.1109/iembs.1998.746084 · Cited by 3
In situ dynamic control of neurite growth by femtosecond laser ablation of substrate patterns
Okano, K., Liu, L.-L., Hosokawa, Y., & Masuhara, H. (2016) · Micro-NanoMechatronics and Human Science · doi:10.1109/mhs.2016.7824186 · Cited by 3
“In vitro construction of a “neuron network on a substrate” that mimics the in vivo architecture and function of a neuropil is expected to facilitate the study of signal processing mechanisms in a brain [1].”
BackgroundBraeken, D., & Prodanov, D. (2010) · InTech eBooks · doi:10.5772/7613 · Cited by 2
“The same group later reported the simultaneous recording of electrical activity and intracellular [Ca2+] using fluorescent dyes, showing the combination of optical and electrical techniques (Jimbo et al. (1993)). www.intechopen.com”
MethodBioassay using living cells integrated on a chip
Torisawa, Y., Shiku, H., Yasukawa, T., & Matsue, T. (2004) · BUNSEKI KAGAKU · doi:10.2116/bunsekikagaku.53.367 · Cited by 5
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“To confine a single neuron within the area of each MEA electrode and form single-cell-per-electrode networks threedimensional microstructures have been used by several researchers [11-13].”
MethodDevelopment of a Neural Interface for PNS Motor Control
Christopher, G., Melinda, K. S., Luo, V. M., Jeffrey, D., & Bonnie, L. F. (2011) · InTech eBooks · doi:10.5772/23896 · Cited by 2
“Generally speaking, these techniques can be divided into two groups based on mechanism: 1) mechanically-based techniques, in which the topography of the electrode itself is in some way modified to cause directed cell adhesion or neurite outgrowth [81, 84] and 2) chemically-based techniques, in which chemical guidance…”
BackgroundMethodMoriguchi, H., Tamai, N., Takayama, Y., & Jimbo, Y. (2007) · International IEEE/EMBS Conference on Neural Engineering · doi:10.1109/cne.2007.369717 · Cited by 2
“In this context, micropatterning[1-10] of cultured”
Self-citationKonno, N., Fukami, T., Shiina, T., & Jimbo, Y. (1998) · doi:10.1541/ieejeiss1987.118.7-8_999 · Cited by 2
Self-citationFrega, M. (2016) · doi:10.1007/978-3-319-30237-9_3 · Cited by 2
Design for Information Processing in Living Neuronal Networks
Kudoh, S. N. (2013) · doi:10.4018/978-1-4666-4225-6.ch003 · Cited by 2
Optimization of thin-film configuration for light-addressable stimulation electrode
Suzurikawa, J., Kanzaki, R., Nakao, M., Jimbo, Y., & Takahashi, H. (2008) · doi:10.1002/ecj.10241 · Cited by 2
Self-citationCoupling of Networks of Neurons to Substrate Planar Microtransducers
Bove, M., Grattarola, M., & Martinoia, S. (1996) · doi:10.1007/978-1-4615-5899-6_20 · Cited by 2
Impact of the entorhinal feed-forward connection to the CA3 on hippocampal coding
Lassers, S. B., Khatri, S. B., Chen, R., Vakilna, Y. S., Tang, W. C., & Brewer, G. J. (2025) · PLoS ONE · doi:10.1371/journal.pone.0326032 · Cited by 1
Fundamental short-term memory of semi-artificial neuronal network
Ito, H., & Kudoh, S. N. (2013) · Annual International Conference of the IEEE Engineering in Medicine and Biology Society · doi:10.1109/embc.2013.6609624 · Cited by 1
“Cultured rat hippocampal neurons form a complex network on a multi-electrodes array (MEA) dish, and spontaneous electrical activity has been observed [1-3].”
BackgroundVitroid - a robot with link between living neuronal network in vitro and robot body
Kudoh, S. N., Tokuda, M., Kiyohara, A., Hosokawa, C., Taguchi, T., & Hayashi, I. (2008) · IEEE International Conference on Mechatronics and Automation (ICMA) · doi:10.1109/icma.2008.4798783 · Cited by 2
“We analysed the spatio-temporal pattern of spontaneous action potentials [3] by multi-site recording system for extracellular potential [4], [6], [5], [3], [7], [8] (Fig.”
MethodSuzurikawa, J., Nakao, M., Kanzaki, R., Jimbo, Y., & Takahashi, H. (2007) · International IEEE/EMBS Conference on Neural Engineering · doi:10.1109/cne.2007.369611 · Cited by 2
Self-citationInteraction and intelligence in living neuronal networks interfaced with moving robot
Kudoh, S. N., & Taguchi, T. (2005) · SPIE Micro + Nano Materials, Devices, and Applications · doi:10.1117/12.638434 · Cited by 1
Characteristics of single neurons cultured on microelectrode arrays in vitro for chemical sensing
Yang, M., Zhang, X., Zhang, Y., & Ozkan, C. (2005) · IEEE Sensors Journal · doi:10.1109/jsen.2005.845520 · Cited by 1
Stochastic frequency signature for chemical sensing using noninvasive neuronelectronic interface
Yang, M., Zhang, X., Zhang, Y., & Ozkan, C. (2005) · IEEE Transactions on Biomedical Engineering · doi:10.1109/tbme.2005.845364 · Cited by 1
“Chip preparation followed previous protocol [22], [23].”
Nam, Y., Wheeler, B. C., & Brewer, G. (2002) · Proceedings of the Second Joint 24th Annual Conference and the Annual Fall Meeting of the Biomedical Engineering Society] [Engineering in Medicine and Biology · doi:10.1109/iembs.2002.1053200 · Cited by 1
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“of axons were emulated along the ditches which were fabricated on an electrode array [ 8 ].”
MethodKasei, M. (1998) · Biofutur · doi:10.1016/s0294-3506(99)80028-3 · Cited by 1
A robust pattern of neuronal response to outer phenomena in "Vitroid", the hybrid neuro-robot
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“The experiment was stopped when Vitroid failed to avoid collision to the wall, or Vitroid reached to the end of the test course.”
BackgroundMoriguchi, H., Tamai, N., Takayama, Y., Kurashima, T., & Jimbo, Y. (2007) · IEEJ Transactions on Electronics Information and Systems · doi:10.1541/ieejeiss.127.1562 · Cited by 1
Self-citationSuzurikawa, J., Takahashi, H., Kanzaki, R., Nakao, M., & Jimbo, Y. (2007) · doi:10.1541/ieejeiss.127.1581 · Cited by 1
Self-citationZhang, Z. D., Yaron, A., Akita, D., Shiramatsu, T., Chao, Z. C., & Takahashi, H. (2025) · European Journal of Neuroscience · doi:10.1111/ejn.70706
“…Scientific), horse serum (Cytiva, Marlborough, MA, USA), GlutaMAX (Thermo Fisher Scientific), and sodium pyruvate (Thermo Fisher Scientific), as described previously (Jimbo et al. 1993; Potter and DeMarse 2001; Yada et al. 2016, 2017; Kubota et al. 2019; Ikeda et al. 2025; Zhang et al. 2025).”
Zhai, X.-J., Jaber, F., Bensaali, F., & Mishra, A. (2020) · doi:10.5013/ijssst.a.16.03.04
Does Representation of Outer Objects in Living Neuronal Network Synthesize “the concept”?
Shibata, E., Ito, H., Minoshima, W., & Kudoh, S. N. (2016) · 2016 Joint 8th International Conference on Soft Computing and Intelligent Systems (SCIS) and 17th International Symposium on Advanced Intelligent Systems (ISIS) · doi:10.1109/scis-isis.2016.0125
Validation of long-term changes of evoked response with self-orgnization map
Fukui, Y., Ito, H., Minoshima, W., & Kudoh, S. N. (2014) · Joint International Conference on Soft Computing and Intelligent Systems and International Symposium on Advanced Intelligent Systems (SCIS&ISIS) · doi:10.1109/scis-isis.2014.7044829 · Cited by 1
Discussion about “synthetic intelligence” in dissociated culture system
Kudoh, S. N., Kiyohara, A., & Taguchi, T. (2011) · doi:10.1002/ecj.10328
Yasuda, K., Kaneko, T., & Nomura, F. (2010) · doi:10.2174/22102892010010100168
MICROPATTERNED POLYMER STRUCTURES FOR CELL AND TISSUE ENGINEERING
Jang, K.-J., Kim, D.-H., Kim, S. M., Levchenko, A., & Suh, K. (2010) · doi:10.1142/9789814295680_0005
Kudoh, S. N., Kiyohara, A., Hosokawa, C., Taguchi, T., & Hayashi, I. (2007) · International Symposium on Micro-NanoMechatronics and Human Science · doi:10.1109/mhs.2007.4420824 · Cited by 1
“Using rat hippocampal cells cultured on multi-electrode arrays[4], [7], [13], we investigated the spatio-temporal pattern of spontaneous action potentials [8].”
BackgroundCholinergic effects on spectral properties of spike trains in rat cortical neurons
Tateno, T., Jimbo, Y., & Robinson, H. P. C. (2004) · 2004 IEEE International Joint Conference on Neural Networks (IEEE Cat. No.04CH37541) · doi:10.1109/ijcnn.2004.1381192
Self-citationIto, Y., Yagi, T., Kanda, H., Tanaka, S., Watanabe, M., & Uchikawa, Y. (1999) · IJCNN'99. International Joint Conference on Neural Networks. Proceedings (Cat. No.99CH36339) · doi:10.1109/ijcnn.1999.831480
“It is known that the axons are guided with various ways such as stereotropism, contact-mediated repulsion, galvanotropism, and so on. For example, some conventional works showed that, the extensions of axons were controlled along ditches which were fabricated on an electrode array [ 7 ].”
BackgroundGrattarola, M., Bove, M., Varreschi, G., & Tedesco, M. (1996) · Proceedings of 18th Annual International Conference of the IEEE Engineering in Medicine and Biology Society · doi:10.1109/iembs.1996.656972
Synchronous bursting and asynchronous firing activity in cultured cortical neurons
Bove, M., Maeda, E., Canepari, M., Grattarola, M., & Kawana, A. (1996) · Proceedings of 18th Annual International Conference of the IEEE Engineering in Medicine and Biology Society · doi:10.1109/iembs.1996.656971
Self-citationDevelopmental Process and Synchronized Bursts in Cultured Neuronal Networks
Ito, D., & Gohara, K. (2014) · Seibutsu Butsuri · doi:10.2142/biophys.54.210
工藤, 卓., 清原, 藍., & 田口, 隆. (2009) · doi:10.1541/ieejeiss.129.32
17 – Controlled cell deposition techniques
Mason, C. (2005) · Elsevier eBooks · doi:10.1533/9781845690809.3.465
Micro and nanopatterned biodegradable polymer films for peripheral nerve regeneration
Miller, C. A. (2000) · doi:10.31274/rtd-180813-767
“, 1988; Kawana, 1996), and silica glass (Jimbo et al., 1993) have been successfully used to study the physical guidance responses of neurons.”
BackgroundJimbo, Y. (1999) · The Brain & Neural Networks · doi:10.3902/jnns.6.24
Self-citationMulti-Site Recording of Neural Activity using Planar Electrode Arrays
Jimbo, Y., & Kawana, A. (1998) · doi:10.1007/978-3-642-58955-3_9
Self-citation
Metadata
- Authors
- Jimbo, Y., Robinson, H. P. C., & Kawana, A.
- Affiliations
- NTT (Japan) (JP)
- Venue
- IEEE Transactions on Biomedical Engineering
- Published
- 1993
- Volume
- 40
- Issue
- 8
- Pages
- 804-810
- ISSN
- 0018-9294 (ISSN-L)
- OpenAlex
- W2051002031
- Cited by
- 192
- References
- 0
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 2 · 2021: 3 · 2020: 2 · 2019: 3 · 2018: 2 · 2017: 1 · 2016: 7 · 2015: 1 · 2014: 5 · 2013: 8 · 2012: 3
- FWCI
- 1.43
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- IEEE Transactions on Biomedical Engineering (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neural Engineering
- Life Sciences › Neuroscience › Cognitive Neuroscience › Neural dynamics and brain function
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- intracellular calcium, microelectrode arrays, fluo-3, cortical neurons, neuronal bursting, electrical stimulation, cell positioning, synaptic connectivity, simultaneous recording
- MeSH terms
- Animals, Calcium, Cells, Cultured, Cerebral Cortex, Cytological Techniques, Electric Stimulation, Electrophysiology, Embryo, Mammalian, Equipment Design, Magnesium, Microelectrodes, Nerve Net, Neurons, Semiconductors, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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