NINscope, a versatile miniscope for multi-region circuit investigations.

Andres De Groot,Romano M Van Genderen,John Van Veldhuijzen,Bastijn Jg Van Den Boom,Chris I De Zeeuw,Joop Bos,Joris Coppens,Ingo Willuhn,Hugo Hoedemaker,Mario Negrello,Tycho M Hoogland

doi:10.7554/elife.49987

Abstract

Miniaturized fluorescence microscopes (miniscopes) have been instrumental to monitor neural signals during unrestrained behavior and their open-source versions have made them affordable. Often, the footprint and weight of open-source miniscopes is sacrificed for added functionality. Here, we present NINscope: a light-weight miniscope with a small footprint that integrates a high-sensitivity image sensor, an inertial measurement unit and an LED driver for an external optogenetic probe. We use it to perform the first concurrent cellular resolution recordings from cerebellum and cerebral cortex in unrestrained mice, demonstrate its optogenetic stimulation capabilities to examine cerebello-cerebral or cortico-striatal connectivity, and replicate findings of action encoding in dorsal striatum. In combination with cross-platform acquisition and control software, our miniscope is a versatile addition to the expanding tool chest of open-source miniscopes that will increase access to multi-region circuit investigations during unrestrained behavior.

Highlights

Cellular resolution imaging using miniaturized fluorescence microscopes permits the monitoring of the topology of activity in brain circuits during unrestrained behaviors
To allow optogenetic stimulation remote from the site of imaging, we developed a simple LED probe consisting of a Mill-Max connector, insulated enamel wires and an SMD LED with 402 case size, which was sealed with epoxy (Figure 1D)
Using the integrated optogenetic LED driver of NINscope we demonstrate its use by stimulating cerebellar Purkinje cells in transgenic Pcp2-Cre Jdhu x Ai32(RCL-ChR2(H134R)/EYFP) mice, while performing calcium imaging from neurons in motor cortex transduced with GCaMP6f (Figure 4A–B, GRIN objective center AP: +1.4, ML: 1.5)

Summary

Introduction

Cellular resolution imaging using miniaturized fluorescence microscopes (miniscopes) permits the monitoring of the topology of activity in brain circuits during unrestrained behaviors. Imaging approaches can reveal how individual cells embedded in a larger network display coordinated activity during different stages of behavior or training (Wagner et al, 2017; Heffley et al, 2018; Galinanes et al, 2018; Giovannucci et al, 2017; Kostadinov et al, 2019) Because of their ability to record in freely moving animals, miniscopes have been instrumental in uncovering neural activity patterns occurring during natural behaviors and related brain-states including social interactions (Murugan et al, 2017; Remedios et al, 2017; Liang et al, 2018; Kingsbury et al, 2019) or sleep (Chen et al, 2018; Cox et al, 2016) with fully intact vestibular input. NINscope is an open-source project enabling others to build on its design and functionalities, thereby contributing to a growing range of opensource tools to study neural circuits during unrestrained behavior

Results

Discussion

Materials and methods

Funding Funder