Chronic, cortex-wide imaging of specific cell populations during behavior

Abstract

Measurements of neuronal activity across brain areas are important for understanding the neural correlates of cognitive and motor processes like attention, decision-making, and action selection. However, techniques that allow cellular resolution measurements are expensive and require a high degree of technical expertise, which limits their broad use. Widefield imaging of genetically encoded indicators is a high throughput, cost effective, and flexible approach to measure activity of specific cell populations with high temporal resolution and a cortex-wide field of view. Here we outline our protocol for assembling a widefield setup, a surgical preparation to image through the intact skull, and imaging neural activity chronically in behaving, transgenic mice that express a calcium indicator in specific subpopulations of cortical neurons. Further, we highlight a processing pipeline that leverages novel, cloud-based methods to analyze large-scale imaging datasets. The protocol targets labs that are seeking to build macroscopes, optimize surgical procedures for long-term chronic imaging, and/or analyze cortex-wide neuronal recordings.