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

Elife. 2017 Sep 25:6:e29839. doi: 10.7554/eLife.29839.

Abstract

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.

Keywords: electrophysiology; medicinal leech; motor control; neuroscience; sensory processing; voltage sensitive dye; whole-brain imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials
  • Animals
  • Behavior, Animal
  • Ganglia, Invertebrate / physiology*
  • Leeches / physiology*
  • Membrane Potentials*
  • Microscopy / methods*
  • Sensation
  • Voltage-Sensitive Dye Imaging / methods*