Abstract
Fluorescence microendoscopy is becoming a promising approach for deep brain imaging, but the current technology for visualizing neurons on a single focal plane limits the experimental efficiency and the pursuit of three-dimensional functional neural circuit architectures. Here we present a novel fast varifocal two-photon microendoscope system equipped with a gradient refractive index (GRIN) lens and an electrically tunable lens (ETL). This microendoscope enables quasi-simultaneous imaging of the neuronal network activity of deep brain areas at multiple focal planes separated by 85-120 µm at a fast scan rate of 7.5-15 frames per second per plane, as demonstrated in calcium imaging of the mouse dorsal CA1 hippocampus and amygdala in vivo.
Highlights
Integrative brain functions such as sensation, perception, cognition, and movement are supported by distributed neuronal networks across different brain areas
Fluorescence microendoscopy is becoming a promising approach for deep brain imaging, but the current technology for visualizing neurons on a single focal plane limits the experimental efficiency and the pursuit of three-dimensional functional neural circuit architectures
We present a novel fast varifocal two-photon microendoscope system equipped with a gradient refractive index (GRIN) lens and an electrically tunable lens (ETL)
Summary
Integrative brain functions such as sensation, perception, cognition, and movement are supported by distributed neuronal networks across different brain areas. Since its introduction into neuroscience in the 1990s, two-photon excitation laser-scanning microscopy [1, 2] has greatly advanced our understanding of the cellular and local circuit bases for these brain functions by visualizing the structure and function of neurons with high resolution in the living brain, the cerebral cortex. There is a technical limitation in that a typical two-photon microscope can image only those neurons that lie within a 1-mm depth in vivo. This has prevented us from gaining a more complete picture of how the entire brain works. Expanding the repertoire of techniques for optical monitoring of deep brain activity would push back the frontiers of neuroscience
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