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Updated: Oct 13, 2025

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Published on: July 24, 2021

Large-scale voltage imaging in behaving mice using targeted illumination.

Sheng Xiao¹, Eric Lowet¹, Howard J Gritton^1,2

¹Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.

|November 11, 2021

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This study introduces targeted illumination to improve widefield voltage imaging of neurons. The new method enhances signal contrast and reduces background noise for clearer optical imaging in behaving animals.

Area of Science:

Neuroscience
Optical Imaging
Biophysics

Background:

Genetically encoded voltage indicators allow optical imaging of neural activity.
Widefield microscopy is crucial for high-speed, large-area neural imaging but suffers from background contamination.
Existing methods lack optical sectioning, limiting signal clarity.

Purpose of the Study:

To develop and validate a targeted illumination strategy for widefield voltage imaging.
To improve signal-to-noise ratio and reduce artifacts in optical recordings.
To enable simultaneous imaging of numerous neurons over large areas in behaving animals.

Main Methods:

Implementation of a digital-micromirror-device (DMD)-based targeted illumination system.
Experimental validation using SomArchon fluorescent voltage indicator in neurons.

Keywords:

Neuroscience Optical imaging Techniques in neuroscience

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Last Updated: Oct 13, 2025

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Quantitative analysis of signal contrast, photobleaching, and background cross-contamination.

Main Results:

Targeted illumination significantly increased SomArchon signal contrast.
The strategy effectively reduced background cross-contamination.
Photobleaching was decreased, allowing for longer recording durations.
Simultaneous imaging of tens of spiking neurons was achieved over minutes in behaving mice.

Conclusions:

DMD-based targeted illumination is a simple yet effective solution for widefield voltage imaging.
This approach overcomes limitations of traditional widefield microscopy for large-scale neural recordings.
The method facilitates high-resolution, large-area optical imaging of neural activity in vivo.