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Digital Scanning Lightsheet Microscopy (DSLM) for Drosophila Whole Brain Functional Imaging
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Digital Scanning Lightsheet Microscopy (DSLM) for Drosophila Whole Brain Functional Imaging

H. Chang, Z.-Y. Chang, K.-C. Jhan, C.-C. Lo, S.-C. Wu, B.-C. Chen 和 L.-A. Chu
Progress in Biomedical Optics and Imaging - Proceedings of SPIE, 卷.13303
2025
Web of Science ID: WOS:001492789600005

摘要

Bessel beam Drosophila Melanogaster Lightsheet microscopy Neuron calcium imaging Odor preference and behavior Olfactory system Confocal microscopy Functional neuroimaging Optical design Structural dynamics Bessel beam Calcium imaging Digital scanning Drosophilla Drosophilla melanogaster Functional imaging Light-sheet microscopies Neuron calcium imaging Odor preference and behavior Olfactory systems Bessel functions
Intravital imaging has presented a powerful tool for tracking cellular function and structural dynamics in the real environment of a complex multicellular organism. However, most commercial microscopy systems are limited by optical design, allowing only long-term live imaging of transparent samples, such as Drosophila embryo, zebrafish embryo, and suffering from slow image acquisition speeds. Consequently, they can only generate two-dimensional time-lapse images at selected optical planes. Here, we introduce a high-resolution Bessel beam based vertical digital scanning light sheet microscopy (V-SPIM) for real-time imaging of the adult fruit fly brain. Compared to traditional multiphoton imaging or multi-point confocal microscopy, Bessel beam based light sheet microscope exhibits lower photodamage and faster acquisition volume rates (V.R.) higher than 1Hz (with proper exposure time (E.T.) and signal to background ratio (SNR)) for each volume(330×330×162μm), allowing us to perform long-term volumetric time-lapse functional imaging in adult animals. This novel approach allows us to visualize the olfactory coding of various odors in a multitude of cells within a single fly brain. © 2025 SPIE.

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