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Published October 30, 2018 | Supplemental Material + Submitted
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Whole-ganglion imaging of voltage in the medicinal leech using a double-sided microscope


Studies of neuronal network emergence during sensory processing and motor control are greatly promoted 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 Hirudo verbana. 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

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. bioRxiv preprint first posted online Jun. 2, 2017. 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 authors declare no competing interests.

Attached Files

Submitted - 145144.full.pdf

Supplemental Material - 145144-1.pdf

Supplemental Material - 145144-2.mov

Supplemental Material - 145144-3.mov

Supplemental Material - 145144-4.mov

Supplemental Material - 145144-5.mov


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August 19, 2023
October 18, 2023