Photometric Patch Electrode to Simultaneously Measure Neural Electrical Activity and Optical Signal in the Brain Tissue

  • Harunori OhmoriEmail author
Part of the Neuromethods book series (NM, volume 152)


Photometric patch electrode (PME) measures optical signal of neurons simultaneously with electrical activity in deep brain tissues. As a light guide, PME transmits laser light to the tip of electrode to excite fluorophores within neurons. The emitted fluorescence from these neurons is captured by the PME simultaneously with the electrical activity. The optical signal is further transmitted through an optical fiber bundle to light detectors; either a photomultiplier tube or a spectrometer. The photomultiplier tube is used for a high-speed monitoring of fluorescence signal in a time range, while the spectrometer is used to analyze changes of a fluorescence signal-profile in a wavelength range. Captured electrical signal and fluorescence signal are highly correlated both in time course and amplitude. Furthermore, PME can apply chemicals locally in the brain tissue by pressure-control within the electrode. We will describe in detail the fabrication of PME and individual components of the PME recording system and demonstrate the application of PME in vitro in brain slices or in vivo in the brain tissue. As one of the limitations of using PME in in vivo experiments, difficulties of labeling neurons by calcium indicators in deep brain tissues are discussed.

Key words

Photometry Patch electrode Calcium response Spectrometer Photomultiplier tube 



We appreciate Dr. Eri Nishino who conceived the experiment together with H.O., and Y. Hirai who improved the software and conducted most experiments using PMT. We thank Drs. R. Matsui and D. Watanabe in Kyoto University for providing us A3V that encodes mCherry and GCaMP6. Experiments in the mouse hippocampus were conducted together with Dr. M. Ono in Kanazawa Medical University. This work was supported by Grants-in-Aid from Japan Society for the Promotion of Science to E. Nishino (23650205) and H. O. (20220008 and 26560464).


  1. 1.
    Berridge MJ, Lipp P, Bootman MD (2000) The versatility and universality of calcium signaling. Nat Rev 1:11–21CrossRefGoogle Scholar
  2. 2.
    Adelsberger H, Garaschuk O, Konnerth A (2005) Cortical calcium waves in resting newborn mice. Nat Neurosci 8:988–990CrossRefGoogle Scholar
  3. 3.
    Hagenston AM, Bading H (2011) Calcium signaling in synapse-to-nucleus communication. Cold Spring Harb Perspect Biol 3:a004564CrossRefGoogle Scholar
  4. 4.
    Stosiek C, Garaschuk O, Holthoff K, Konnerth A (2003) In vivo two-photon calcium imaging of neuronal networks. Proc Nat Acad Sci U S A 100:7319–7324CrossRefGoogle Scholar
  5. 5.
    Akerboom J, Chen T-W, Wardill TJ et al (2012) Optimization of a GCamp calcium indicato for neural activity imaging. J Neurosci 32:13819–13840PubMedPubMedCentralGoogle Scholar
  6. 6.
    Ikegaya Y, Aaron G, Cossart R et al (2004) Synfire chains and cortical songs: temporal modules of cortical activity. Science 304:559–564CrossRefGoogle Scholar
  7. 7.
    Ohki K, Chung S, Ch’ng YH, Kara P, Reid RC (2005) Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex. Nature 433:597–603CrossRefGoogle Scholar
  8. 8.
    Helmchen F, Denk W (2005) Deep tissue two-photon microscopy. Nat Methods 2:932–940CrossRefGoogle Scholar
  9. 9.
    Svoboda K, Yasuda R (2006) Principles of two-photon excitation microscopy and its applications to neuroscience. Neuron 50:823–839CrossRefGoogle Scholar
  10. 10.
    Jung JC, Mehta AD, Aksay E et al (2004) In vivo mammalian brain imaging using one- and two-photon fluorescence microscopy. J Neurophysiol 92:3121–3133CrossRefGoogle Scholar
  11. 11.
    Hayashi Y, Tagawa Y, Yawata S et al (2012) Technical spotlight. Spatio-temporal control of neural activity in vivo using fluorescence microendoscopy. Eur J Neurosci 36:2722–2732CrossRefGoogle Scholar
  12. 12.
    Vincent P, Makos U, Charvet I et al (2006) Live imaging of neural structure and function by fibered fluorescence microscopy. EMBO Rep 7:1154–1161CrossRefGoogle Scholar
  13. 13.
    LeChasseur Y, Dufour S, Larvertu G et al (2011) A microbe for parallel optical and electrical recordings from single neurons in vivo. Nat Methods 8:319–325CrossRefGoogle Scholar
  14. 14.
    Hirai Y, Nishino E, Ohmori H (2015) Simultaneous recording of fluorescence and electrical signals by photometric patch electrode in deep brain regions in vivo. J Neurophysiol 113:3930–3942CrossRefGoogle Scholar
  15. 15.
    Jarvis E (2005) The Avian Brain Nomenclature Consortium. Avian brains and a new understanding of vertebrate brain evolution. Nat Rev Neurosci 6:151–159CrossRefGoogle Scholar
  16. 16.
    Kawai S, Takagi Y, Kaneko S, Kurosawa T (2011) Effect of three types of mixed anesthetic agents alternate to ketamine in mice. Exp Anim 60:481–487CrossRefGoogle Scholar
  17. 17.
    Fukui I, Sato T, Ohmori H (2006) Improvement of phase information at low sound frequency in nucleus magnocellularis of the chicken. J Neurophysiol 96:633–641CrossRefGoogle Scholar
  18. 18.
    Nishino E, Yamada R, Kuba H et al (2008) Sound-intensity-dependent compensation for the small interaural time difference cue to sound source localization. J Neurosci 28:7153–7164CrossRefGoogle Scholar
  19. 19.
    Murayama M, Miyazaki K, Kudo Y et al (2005) Optical monitoring of progressive synchronization in dentate granule cells during population burst activities. Eur J Neurosci 21:3349–3360CrossRefGoogle Scholar
  20. 20.
    Matsui R, Tanabe Y, Watanabe D (2012) Avian adeno-associated virus vector efficiently transduces neurons in the embryonic and post-embryonic chicken brain. PLoS One 7(11):e48730CrossRefGoogle Scholar
  21. 21.
    Buller RM, Janik JE, Sebring ED, Rose JA (1981) Herpes simplex virus types 1 and 2 completely help adenovirus-associated virus replication. J Virol 40:241–247PubMedPubMedCentralGoogle Scholar
  22. 22.
    McCown TJ, Xiao X, Li J, Breese GR, Samulski RJ (1996) Differential and persistent expression patterns of CNS gene transfer by an adeno-associated virus (AAV) vector. Brain Res 713:99–107CrossRefGoogle Scholar
  23. 23.
    Takatsuka K, Ishii TM, Ohmori H (2005) A novel Ca2+ indicator protein using FRET and calpain-sensitive linker. BBRC 336:316–323PubMedGoogle Scholar
  24. 24.
    Hall MP, Unch J, Binkowski BF et al (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. Chem Biol 7:1848–1857Google Scholar
  25. 25.
    Couturier C, Deprez B (2012) Setting up a bioluminescence resonance energy transfer high throughput screening assay to search for protein/protein interaction inhibitors in mammalian cells. Front Endocrinol 3:100CrossRefGoogle Scholar
  26. 26.
    Emery EC, Luiz AP, Sikandar S et al (2016) In vivo characterization of distinct modality-specific subsets of somatosensory neurons using GCaMP. Sci Adv 2(11):e1600990CrossRefGoogle Scholar
  27. 27.
    Park JE, Zhang XF, Choi S-H et al (2016) Generation of transgenic marmosets expressing genetically encoded calcium indicators. Sci Rep 6:34931CrossRefGoogle Scholar
  28. 28.
    Chen TW, Wardill TJ, Sun Y et al (2013) Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499:295–300CrossRefGoogle Scholar
  29. 29.
    Hendel T, Mank M, Schnell B et al (2008) Fluorescence changes of genetic calcium indicators and OGB-1 correlated with neural activity and calcium in vivo and in vitro. J Neurosci 28(29):7399–7411CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Faculty of Medicine, Department of Neurobiology and PhysiologyKyoto UniversityKyotoJapan

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