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In Vitro Voltage Imaging of Subthreshold Activity in Inferior Olive Neurons with ANNINE-6plus

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Part of the Neuromethods book series (NM,volume 177)

Abstract

Since the 1970s, a multitude of fluorescence-based methods for observing changes in neuronal membrane voltage have been developed, progressively improving in terms of signal sensitivity, speed, and targeting. While significant development has been seen in the past decade in engineering novel variants of genetically encoded voltage indicators (GEVI), synthetic dyes can, in many cases, be more straightforward and flexible tools to be used. Here, we describe in detail an improved dye-labeling method where the synthetic, pure electrochromic voltage-sensitive dye ANNINE-6plus is introduced into the brain region of interest via stereotactical injection in a mouse. Specifically, we have optimized the labeling protocol for effective voltage imaging of subthreshold activity in inferior olive. The homogenous labeling and excellent signal-to-noise ratio allow this method to reveal olivary subthreshold oscillations in high spatiotemporal resolution. In addition to the procedures related to dye introduction and optical imaging, we give a detailed description of the process of acute slice preparation from the mouse inferior olive.

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Correspondence to Marylka Yoe Uusisaari .

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Dorgans, K., Kuhn, B., Uusisaari, M.Y. (2022). In Vitro Voltage Imaging of Subthreshold Activity in Inferior Olive Neurons with ANNINE-6plus. In: Sillitoe, R.V. (eds) Measuring Cerebellar Function. Neuromethods, vol 177. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2026-7_8

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  • DOI: https://doi.org/10.1007/978-1-0716-2026-7_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2025-0

  • Online ISBN: 978-1-0716-2026-7

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