High-resolution focus-tunable one-photon miniaturized fluorescence microscope for imaging in freely moving animals

Xiaoyu Liu; Zenan Wu; Junnan Xu; Wei Gong; Ke Si

doi:10.1117/12.3007740

18 December 2023 High-resolution focus-tunable one-photon miniaturized fluorescence microscope for imaging in freely moving animals

Xiaoyu Liu, Zenan Wu, Junnan Xu, Wei Gong, Ke Si

Author Affiliations +

Proceedings Volume 12963, AOPC 2023: Optical Sensing, Imaging, and Display Technology and Applications; and Biomedical Optics; 129631Z (2023) https://doi.org/10.1117/12.3007740
Event: Applied Optics and Photonics China 2023 (AOPC2023), 2023, Beijing, China

Abstract

The relationship between neural activity, brain function, and corresponding biological behaviors remains a significant challenge in neuroscience. Exploring this relationship needs various optical imaging techniques to acquire real-time data with high spatial resolution. A promising technology recently is wearable miniaturized microscopes (mini scopes), which enable long-term neural activity recording in freely moving animals. However, most one-photon mini-scopes have limitations for imaging in-depth with high resolution and large field of view (FOV). To address this, we developed a one-photon miniaturized fluorescence microscope (1P-miniFM), intended for imaging of live brain neurons in free-behaving animals at subcellular level (~1.2 μm). We conducted specially designed optical path, achieving an imaging FOV of ~700 × 400 μm. In addition, we incorporated an electrowetting lens (EWL) to achieve a wide range of ~300 μm z-axis scanning with little resolution loss. 1P-miniFM is compact (11 × 17 × 24 mm) and lightweight (~2.9 g), causing little impediment to animals’ spontaneous behaviors. With genetically encoded calcium indicator GCaMP6s, we monitored neuron activities in secondary motor cortex (M2) during consecutive pain-related and sensory stimulations. We found that M2 neurons are key components and exhibit distinct variations in the response patterns. 1P-miniFM has potential as an excellent tool to explore relationships between neuron network and animal behaviors.

(2023) Published by SPIE. Downloading of the abstract is permitted for personal use only.

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Xiaoyu Liu, Zenan Wu, Junnan Xu, Wei Gong, and Ke Si "High-resolution focus-tunable one-photon miniaturized fluorescence microscope for imaging in freely moving animals", Proc. SPIE 12963, AOPC 2023: Optical Sensing, Imaging, and Display Technology and Applications; and Biomedical Optics, 129631Z (18 December 2023); https://doi.org/10.1117/12.3007740

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KEYWORDS

Neurons

Animals

Brain

Fluorescence

Image resolution

Biological imaging

Calcium

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