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Published September 25, 2017 | Supplemental Material + Submitted + Published
Journal Article Open

A double-sided microscope to realize whole-ganglion imaging of membrane potential in the medicinal leech


Studies of neuronal network emergence during sensory processing and motor control are greatly facilitated by technologies that allow us to simultaneously record the membrane potential dynamics of a large population of neurons in single cell resolution. To achieve whole-brain recording with the ability to detect both small synaptic potentials and action potentials, we developed a voltage-sensitive dye (VSD) imaging technique based on a double-sided microscope that can image two sides of a nervous system simultaneously. We applied this system to the segmental ganglia of the medicinal leech. Double-sided VSD imaging enabled simultaneous recording of membrane potential events from almost all of the identifiable neurons. Using data obtained from double-sided VSD imaging, we analyzed neuronal dynamics in both sensory processing and generation of behavior and constructed functional maps for identification of neurons contributing to these processes.

Additional Information

© 2017 Tomina and Wagenaar. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: 22 June 2017; Accepted: 25 September 2017; Published: 25 September. We thank Evan Miller for sharing of the VF2.1(OMe).H dye; Annette Stowasser for her role in developing a prototype of the double-sided microscope and many helpful conversations; and Angela Bruno for useful discussions regarding data analysis. This work was supported by the Burroughs Welcome Fund through a Career Award at the Scientific Interface and by the National Institute of Neurological Disorders and Stroke through grant R01 NS094403 (both to DAW). YT was supported by JSPS Overseas Research Fellowships. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Author Contributions: Yusuke Tomina, Conceptualization, Data curation, Software, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing—original draft, Project administration; Daniel A Wagenaar, Conceptualization, Resources, Data curation, Software, Formal analysis, Supervision, Funding acquisition, Validation, Visualization, Methodology, Project administration, Writing—review and editing.

Attached Files

Published - elife-29839.pdf

Submitted - 145144.full.pdf

Supplemental Material - elife-29839-transrepform-v3.docx


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