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Fast confocal fluorescence imaging in freely behaving mice
Clara Dussaux1, Vivien Szabo2, Yan Chastagnier2
1Institut de biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, 46 rue d'Ulm, Paris, 75005, France.
Scientific Reports
|November 4, 2018
Summary
We developed a novel fiberscope for fast, background-free brain fluorescence imaging in mice. This advanced system enables high-resolution visualization of cortical microvasculature and red blood cell velocity in awake animals.
Area of Science:
- Neuroscience
- Biomedical Engineering
- Optical Imaging
Background:
- Miniaturization constraints challenge in vivo brain fluorescence imaging in mice.
- Achieving large field of view, high temporal resolution, and effective background rejection remains difficult.
Purpose of the Study:
- To present a novel fiberscope system for advanced brain fluorescence imaging in freely behaving mice.
- To overcome limitations in field of view, temporal resolution, and background noise.
Main Methods:
- A custom multipoint-scanning confocal microscope coupled to a mouse via an image guide and micro-objective.
- Simultaneous acquisition of confocal and widefield images to subtract out-of-focus background.
- Utilizing a digital micromirror device for adaptable illumination and detection pinholes.
Main Results:
- Fast (up to 200 Hz), background-free fluorescence imaging over a 230 μm field of view.
- Imaging of mouse cortical microvasculature up to 120 μm deep with background reduced by two orders of magnitude.
- Measurement of red blood cell velocity in cortical microvasculature, showing increased velocity in awake vs. anesthetized mice.
Conclusions:
- The novel fiberscope system significantly improves background-free fluorescence imaging in the mouse brain.
- The system enables high-speed imaging for functional studies, such as measuring microvascular blood flow dynamics.
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