Abstract

Although modern techniques such as two-photon microscopy can now provide cellular-level three-dimensional imaging of the intact living brain, the speed and fields of view of these techniques remain limited. Conversely, two-dimensional wide-field optical mapping (WFOM), a simpler technique that uses a camera to observe large areas of the exposed cortex under visible light, can detect changes in both neural activity and haemodynamics at very high speeds. Although WFOM may not provide single-neuron or capillary-level resolution, it is an attractive and accessible approach to imaging large areas of the brain in awake, behaving mammals at speeds fast enough to observe widespread neural firing events, as well as their dynamic coupling to haemodynamics. Although such wide-field optical imaging techniques have a long history, the advent of genetically encoded fluorophores that can report neural activity with high sensitivity, as well as modern technologies such as light emitting diodes and sensitive and high-speed digital cameras have driven renewed interest in WFOM. To facilitate the wider adoption and standardization of WFOM approaches for neuroscience and neurovascular coupling research, we provide here an overview of the basic principles of WFOM, considerations for implementation of wide-field fluorescence imaging of neural activity, spectroscopic analysis and interpretation of results.This article is part of the themed issue ‘Interpreting BOLD: a dialogue between cognitive and cellular neuroscience’.

Highlights

  • Wide-field optical imaging of neural activity was first demonstrated 30 years ago using cortical application of fluorescent voltage-sensitive dyes (VSDs) [1]

  • Additional studies have demonstrated that the dynamics of flavoprotein fluorescence can be optically mapped in the brain of wild-type animals as an indicator of oxidative metabolism [18,19]

  • These data were processed using methods for converting reflectance data to haemodynamic parameters, while fluorescence recordings were corrected to compensate for haemodynamic cross-talk

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Summary

Introduction

Wide-field optical imaging of neural activity was first demonstrated 30 years ago using cortical application of fluorescent voltage-sensitive dyes (VSDs) [1]. Additional studies have demonstrated that the dynamics of flavoprotein fluorescence can be optically mapped in the brain of wild-type animals as an indicator of oxidative metabolism [18,19] Such wide-field neuroimaging methods are readily compatible with optogenetic approaches to modulate brain activity in awake animals [20]. These data were processed using methods for converting reflectance data to haemodynamic parameters, while fluorescence recordings were corrected to compensate for haemodynamic cross-talk. The basic principles of both reflectance and fluorescence WFOM are reviewed, including approaches and considerations for analysis of data including conversion of reflectance data into haemodynamic parameters and correction of fluorescence for haemodynamic cross-talk

Reflectance wide-field optical mapping
Wide-field optical mapping of fluorescence contrast
Summary
58. Hall CN et al 2014 Capillary pericytes regulate
57. Tian P et al 2010 Cortical depth-specific
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