Formation of essential ultrastructural interface between cultured hippocampal cells and gold mushroom-shaped MEA- towards "IN-CELL" recordings from vertebrate neurons

A. Fendyur, N. Mazurski, J. Shappir, M.E. Spira

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


Using cultured Aplysia neurons we recently reported on the development of a novel approach in which an extracellular, noninvasive multi-electrode-array system provides multisite, attenuated, intracellular recordings of sub-threshold synaptic potentials and action potentials (APs), the so called "IN-CELL" recording configuration. Because of its noninvasive nature, the configuration can be used for long term semi intracellular electrophysiological monitoring of APs and synaptic potentials. Three principals converge to generate the IN-CELL configuration: (a) engulfment of approximately one micrometer size gold mushroom-shaped microelectrodes (gMμE) by the neurons, (b) formation of high seal resistance between the cell's plasma membrane and the engulfed gMμE and (c), autonomous localized increased conductance of the membrane patch facing the gMμE. Using dissociated rat hippocampal cultures we report here that the necessary morphological and ultrastructural relationships to generate the IN-CELL recording configuration are formed between hippocampal cells and the gMμEs. Interestingly, even
Original languageEnglish
JournalFrontiers in Neuroengineering
Issue numberNOVEMBER
StatePublished - 2011

Bibliographical note

Cited By :38

Export Date: 11 September 2022

Correspondence Address: Spira, M. E.; Department of Neurobiology, , Jerusalem, 91940, Israel; email:

References: Akaike, N., Harata, N., Nystatin perforated patch recording and its applications to analyses of intracellular mechanisms (1994) Jpn J Physiol,, 44, pp. 433-473; Ambros-Ingerson, J., Holmes, W.R., Analysis and comparison of morphological reconstructions of hippocampal field CA1 pyramidal cells (2005) Hippocampus,, 15, pp. 302-315; Berdondini, L., Massobrio, P., Chiappalone, M., Tedesco, M., Imfeld, K., McCione, A., Gandolfo, M., Martinoia, S., Extracellular recordings from locally dense microelectrode arrays coupled to dissociated cortical cultures (2009) J Neurosci Methods,, 177, pp. 386-396; Claverol-Tinture, E., Pine, J., Extracellular potentials in low-density dissociated neuronal cultures (2002) J Neurosci Meth, 117, pp. 13-21; Cohen, A., Shappir, J., Yitzchaik, S., Spira, M.E., Reversible transition of extracellular field potential recordings to intracellular recordings of action potentials generated by neurons grown on transistors (2008) Biosens Bioelectron,, 23, pp. 811-819; Craig, A.M., Banker, G., Neuronal polarity (1994) Annu Rev Neurosci,, 17, pp. 267-310; Einevoll, G.T., Wojcik, D.K., Destexhe, A., Modeling extracellular potentials (2010) J Comput Neurosci,, 29, pp. 367-369; Fromherz, P., (2003) Neuroelectronic interfacing: Semiconductor chips with ion channels, nerve cells, and brain., , Berlin: Wiley-VCH; Fromherz, P., Three levels of neuroelectronic interfacing: Silicon chips with ion channels, nerve cells, and brain tissue (2006) Ann N Y Acad Sci,, 1093, pp. 143-160; Gitler, D., Xu, Y., Kao, H.T., Lin, D., Lim, S., Feng, J., Greengard, P., Augustine, G.J., Molecular determinants of synapsin targeting to presynaptic terminals (2004) J Neurosci,, 24, pp. 3711-3720; Hai, A., Dormann, A., Shappir, J., Yitzchaik, S., Bartic, C., Borghs, G., Langedijk, J.P., Spira, M.E., Spine-shaped gold protrusions improve the adherence and electrical coupling of neurons with the surface of micro-electronic devices (2009) J R Soc Interface,, 6, pp. 1153-1165; Hai, A., Kamber, D., Malkinson, G., Erez, H., Mazurski, N., Shappir, J., Spira, M.E., Changing gears from chemical adhesion of cells to flat substrata toward engulfment of micro-protrusions by active mechanisms (2009) J Neural Eng,, 6, p. 066009; Hai, A., Shappir, J., Spira, M.E., In-cell recordings by extracellular microelectrodes (2010) Nat Methods,, 7, pp. 200-202; Hai, A., Shappir, J., Spira, M.E., Long-term, multisite, parallel, in-cell recording and stimulation by an array of extracellular microelectrodes (2010) J Neurophysiol,, 104, pp. 559-568; Halassa, M.M., Haydon, P.G., Integrated brain circuits: Astrocytic networks modulate neuronal activity and behavior (2010) Annu Rev Physiol,, 72, pp. 335-355; Hochberg, L.R., Serruya, M.D., Friehs, G.M., Mukand, J.A., Saleh, M., Caplan, A.H., Branner, A., Donoghue, J.P., Neuronal ensemble control of prosthetic devices by a human with tetraplegia (2006) Nature,, 442, pp. 164-171; Inyushin, M., Tsytsarev, V., Ignashchenkova, A., Lenkov, D.N., Use of artificial ion channels for quasi-intracellular recording of cerebral cortex neuron activity (1997) Neurosci Behav Physiol,, 27, pp. 702-707; Jenkner, M., Fromherz, P., Bistability of membrane conductance in cell adhesion observed in a neuron transistor (1997) Phys. Rev. Lett.,, 79, pp. 4705-4708; Johnstone, A.F., Gross, G.W., Weiss, D.G., Schroeder, O.H., Gramowski, A., Shafer, T.J., Microelectrode arrays: A physiologically based neurotoxicity testing platform for the 21st century (2010) Neurotoxicology,, 31, pp. 331-350; Lebedev, M.A., Nicolelis, M.A., Brain-machine interfaces: Past, present and future (2006) Trends Neurosci,, 29, pp. 536-546; Linden, D.J., Long-term potentiation of glial synaptic currents in cerebellar culture (1997) Neuron,, 18, pp. 983-994; McCione, A., Gandolfo, M., Tedesco, M., Nieus, T., Imfeld, K., Martinoia, S., Berdondini, L., Experimental Investigation on Spontaneously Active Hippocampal Cultures Recorded by Means of High-Density MEAs: Analysis of the Spatial Resolution Effects (2010) Front Neuroeng,, 10, pp. 3-4; Markram, H., Perin, R., Innate neural assemblies for lego memory (2011) Front Neural Circuits,, 5, p. 6; McAdams, E.T., Jossinet, J., Subramanian, R., McCauley, R.G., Characterization of gold electrodes in phosphate buffered saline solution by impedance and noise measurements for biological applications (2006) Conf Proc IEEE Eng Med Biol Soc,, 1, pp. 4594-4597; McMahon, H.T., Gallop, J.L., Membrane curvature and mechanisms of dynamic cell membrane remodelling (2005) Nature,, 438, pp. 590-596; Mennerick, S., Benz, A., Zorumski, C.F., Components of glial responses to exogenous and synaptic glutamate in rat hippocampal microcultures (1996) J Neurosci,, 16, pp. 55-64; Mennerick, S., Zorumski, C.F., Glial contributions to excitatory neurotransmission in cultured hippocampal cells (1994) Nature,, 368, pp. 59-62; Mirsky, V.M., Riepl, M., Wolfbeis, O.S., Capacitive monitoring of protein immobilization and antigen-antibody reactions on monomolecular alkylthiol films on gold electrodes (1997) Biosens Bioelectron,, 12, pp. 977-989; Murphy, T.H., Blatter, L.A., Wier, W.G., Baraban, J.M., Rapid communication between neurons and astrocytes in primary cortical cultures (1993) J Neurosci,, 13, pp. 2672-2679; Nam, Y., Branch, D.W., Wheeler, B.C., Epoxy-silane linking of biomolecules is simple and effective for patterning neuronal cultures (2006) Biosens Bioelectron,, 22, pp. 589-597; Orkand, R.K., Nicholls, J.G., Kuffler, S.W., Effect of nerve impulses on the membrane potential of glial cells in the central nervous system of amphibia (1966) J Neurophysiol,, 29, pp. 788-806; Perin, R., Berger, T.K., Markram, H., A synaptic organizing principle for cortical neuronal groups (2011) Proc Natl Acad Sci,, 108, pp. 5419-5424; Quiroga, R.Q., Nadasdy, Z., Ben-Shaul, Y., Unsupervised spike detection and sorting with wavelets and superparamagnetic clustering (2004) Neural Comput,, 16, pp. 1661-1687; Rothman, S., Cowan, W.M., A scanning electron microscope study of the in vitro development of dissociated hippocampal cells (1981) J Comp Neurol,, 195, pp. 141-155; Sakmann, B., Neher, E., Patch clamp techniques for studying ionic channels in excitable membranes (1984) Annu Rev Physiol,, 46, pp. 455-472; Schipke, C.G., Kettenmann, H., Astrocyte responses to neuronal activity (2004) Glia,, 47, pp. 226-232; Scorza, C.A., Araujo, B.H., Leite, L.A., Torres, L.B., Otalora, L.F., Oliveira, M.S., Garrido-Sanabria, E.R., Cavalheiro, E.A., Morphological and electrophysiological properties of pyramidal-like neurons in the stratum oriens of Cornu ammonis 1 and Cornu ammonis 2 area of Proechimys (2011) Neuroscience,, 177, pp. 252-268; Soussou, W.V., Yoon, G.J., Brinton, R.D., Berger, T.W., Neuronal network morphology and electrophysiologyof hippocampal neurons cultured on surface-treated multielectrode arrays (2007) IEEE Trans Biomed Eng,, 54, pp. 1309-1320; Spira, M.E., Kamber, D., Dormann, A., Cohen, A., Bartic, C., Borghs, G., Langedijk, J.P.M., Shappir, J., Improved neuronal adhesion to the surface of electronic device by engulfment of protruding micro-nails fabricated on the chip surface (2007) Transducers '07 & Eurosensors Xxi Digest of Technical Papers, 1-2, pp. U628; Spira, M.E., Oren, R., Dormann, A., Gitler, D., Critical calpain-dependent ultrastructural alterations underlie the transformation of an axonal segment into a growth cone after axotomy of cultured Aplysia neurons (2003) J Comp Neurol,, 457, pp. 293-312; Spruston, N., Johnston, D., Perforated patch-clamp analysis of the passive membrane properties of three classes of hippocampal neurons (1992) J Neurophysiol,, 67, pp. 508-529; Studer, D., Humbel, B.M., Chiquet, M., Electron microscopy of high pressure frozen samples: Bridging the gap between cellular ultrastructure and atomic resolution (2008) Histochem Cell Biol,, 130, pp. 877-889; Velez-Fort, M., Audinat, E., Angulo, M.C., Central Role of GABA in Neuron-Glia Interactions (2011) Neuroscientist.; Wheeler, B.C., Nam, Y., In vitro microelectrode array technology and neural recordings (2011) Crit Rev Biomed Eng,, 39, pp. 45-61


  • Action potential
  • Electrical coupling
  • Field potentials
  • Gold mushroom-shaped microelectrodes
  • MEA
  • Multi-electrode-array
  • animal cell
  • article
  • brain computer interface
  • cell activity
  • cell membrane
  • cell structure
  • controlled study
  • electroencephalogram
  • gold mushroom shape microelectrode
  • hippocampus
  • information processing
  • intracellular recording
  • microelectrode
  • nerve cell
  • nerve potential
  • nervous system conductance
  • nervous system electrophysiology
  • newborn
  • nonhuman
  • rat
  • synaptic potential


Dive into the research topics of 'Formation of essential ultrastructural interface between cultured hippocampal cells and gold mushroom-shaped MEA- towards "IN-CELL" recordings from vertebrate neurons'. Together they form a unique fingerprint.

Cite this