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

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

Abstract

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 

Notes

Acknowledgments

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).

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Copyright information

© 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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