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20 August 2020 Light sheet microscopy for fast functional imaging of 3D neuronal cultures in hydrogels
Gustavo Castro-Olvera, Jorge Madrid-Wolff, Omar E. Olarte, Estefanía Estévez-Priego, Adriaan A. Ludl, Emilio J. Gualda, Laurent Ladepeche, Jordi Soriano, Pablo Loza-Alvarez
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Proceedings Volume 11469, Emerging Topics in Artificial Intelligence 2020; 114690C (2020) https://doi.org/10.1117/12.2567263
Event: SPIE Nanoscience + Engineering, 2020, Online Only
Abstract
I will present a Light-sheet fluorescence microscope (LSFM) for fast volumetric imaging during extended periods of time. In this case, the observation arm of the microscope contains an electrically tunable lens (ETL) that is used to shift the focal position of the collection lens. By moving the light sheet plane in synchrony with the ETL, it is possible to scan the full 3D sample, which remains totally static, at high speeds (25 volumes/s) [2]. This system is used to image the spontaneous Ca2+ activity, as reported by GCaMP fluorescence, in 3D of primary neuron cultures in hydrogels. The field of view is of 300µm x 300µm x 1mm. The imaging speeds allows a proper sampling of the propagation of GCaMP signal in the full observation volume [4]. The obtained data is then processed to calculate the connectivity maps in the 3D neuron cultured in hydrogels.
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Gustavo Castro-Olvera, Jorge Madrid-Wolff, Omar E. Olarte, Estefanía Estévez-Priego, Adriaan A. Ludl, Emilio J. Gualda, Laurent Ladepeche, Jordi Soriano, and Pablo Loza-Alvarez "Light sheet microscopy for fast functional imaging of 3D neuronal cultures in hydrogels", Proc. SPIE 11469, Emerging Topics in Artificial Intelligence 2020, 114690C (20 August 2020); https://doi.org/10.1117/12.2567263
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KEYWORDS
Microscopy
3D image processing
Functional imaging
Luminescence
Microscopes
Neurons
3D scanning
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