Real-Time Calcium Imaging of a Mouse Medial Prefrontal Cortex Using a Miniscope

Abstract

Source: Thapa, R., et. al., Stereotaxic Viral Injection and Gradient-Index Lens Implantation for Deep Brain In Vivo Calcium Imaging. J. Vis. Exp. (2021)

The video demonstrates real-time calcium imaging using a miniscope in a transduced mouse, correlating neuronal activity with behavior.

Protocol

Miniscope mounting and in vivo Ca²⁺ imaging (Figure 1)

1. Mount the miniscope to its base
   1. Anesthetize the mouse briefly in the induction chamber (5% isoflurane and 1 L/min oxygen flow rate). Place the mouse on a clean bench surface.
   2. Loosen the locking screw with a small screwdriver, remove the protective cap, and clean the surface of the GRIN (Gradient-index lens) lens with an acetone-soaked cotton swab.
   3. Wrap a PTFE tape (Polytetrafluoroethylene tape) tightly around the miniscope thread and fasten the miniscope to its base on the mouse head.....

Results

Figure 1: Schematic overview of the experimental procedure. (A.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Acetone	Sigma-Aldrich	179124-1L	GRIN lens cleaner
Behavior Recoding Software (Point Grey FlyCap2)	Point Grey	Point Grey Research Blackfly BFLY-PGE-12A2C	For recording behavior
Data acquisition controller	NIDA/IRP	Custom-designed	Component of image acquisition system
Data transmission cable	NIDA/IRP	Custom-designed	Component of image acquisition system
GRIN lens	GRINTECH	NEM-100-25-10-860-S	For GRIN lens implantation
Miniscope base	Protolabs	Custom-designed	For mounting the base
Miniscope holding arm	NIDA/IRP	Custom-designed	For mounting the base
Miniscope protection cap	Protolabs	Custom-designed	For protecting the miniscope
Motorized controller	Thorlabs	KMTS50E	For mounting the base
NeuView	NIDA/IRP	https://github.com/giovannibarbera/miniscope_v1.0	For in vivo imaging
Ophthalmic ointment	Puralube Vet Ointment	NDC 17033-211-38	Ophthalmic
Polytetrafluoroethylene (PTFE) tape	TegaSeal PTFE Tape	A-A-58092	For fastening miniScope to the base
Sterile cotton swab	Puritan	REF 806-WC	For both surgeries