A robot for high yield electrophysiology and morphology of single neurons in vivo

Lu Li,William A Stoy,Hongkui Zeng,Christof Koch,Craig R Forest,Emma J Garren,Tanya L Daigle,Benjamin Ouellette

doi:10.1038/ncomms15604

Abstract

Single-cell characterization and perturbation of neurons provides knowledge critical to addressing fundamental neuroscience questions including the structure–function relationship and neuronal cell-type classification. Here we report a robot for efficiently performing in vivo single-cell experiments in deep brain tissues optically difficult to access. This robot automates blind (non-visually guided) single-cell electroporation (SCE) and extracellular electrophysiology, and can be used to characterize neuronal morphological and physiological properties of, and/or manipulate genetic/chemical contents via delivering extraneous materials (for example, genes) into single neurons in vivo. Tested in the mouse brain, our robot successfully reveals the full morphology of single-infragranular neurons recorded in multiple neocortical regions, as well as deep brain structures such as hippocampal CA3, with high efficiency. Our robot thus can greatly facilitate the study of in vivo full morphology and electrophysiology of single neurons in the brain.

Highlights

Single-cell characterization and perturbation of neurons provides knowledge critical to addressing fundamental neuroscience questions including the structure–function relationship and neuronal cell-type classification
Labelling the neurons investigated in vivo allows obtaining and correlating multiple modalities of data including full morphology, function/physiology and/or genetics at the single-neuron level, which is critical to addressing long-standing neuroscience questions such as the structure–function relationship and neuronal cell-type classification[7]
Acquiring and correlating the morphology/connectivity, function/physiology and/or genetics of single neurons will facilitate our understanding of the neuronal structure–function relationship and cell-type classification

Summary

Introduction

Single-cell characterization and perturbation of neurons provides knowledge critical to addressing fundamental neuroscience questions including the structure–function relationship and neuronal cell-type classification. We report a robot for efficiently performing in vivo single-cell experiments in deep brain tissues optically difficult to access This robot automates blind (non-visually guided) single-cell electroporation (SCE) and extracellular electrophysiology, and can be used to characterize neuronal morphological and physiological properties of, and/or manipulate genetic/chemical contents via delivering extraneous materials (for example, genes) into single neurons in vivo. By automating in vivo SCE, ACE can manipulate the chemical and/or genetic contents of single neurons by delivering a variety of extraneous materials into single cells based on the electric charge (electrophoresis) and/or concentration gradient (passive diffusion) through glass micropipettes, the best available option to gain physical access to neurons in deep brain structures. Blind patch recording and SCE have been manually conducted in multiple animal models[23,24,25,26,27,28,29,30], meaning that ACE can have broad applications in various species in which transgenic animal models are not readily available

Methods

Results

Conclusion