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Light Sheet Fluorescence Microscopy (LSFM) is an ideal tool for imaging model organisms that are hundreds of microns in size, providing high resolution and fast frame rates. In order to extend LSFM to rapid 3D volumetric imaging, a popular method is to add an electrically tunable lens (ETL) in the detection path of the LSFM. But for larger fields of view and higher NA objectives, the ETL introduces aberrations in the system. Here, we developed an LSFM with adaptive optics and an ETL for rapid focusing. We demonstrate that the system enables imaging over a volume of 499 × 499 × 140 μm3 with a volumetric speed of 4 Hz. We apply the system to image neural activity in the zebrafish larvae to capture rare seizure events.
Bingxi Liu,Yang Liu,Carly Duffy,James D. Lauderdale, andPeter Kner
"Fast volumetric seizure model imaging in zebrafish brain with light sheet microscopy", Proc. SPIE 12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, 1284808 (12 March 2024); https://doi.org/10.1117/12.3000519
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Bingxi Liu, Yang Liu, Carly Duffy, James D. Lauderdale, Peter Kner, "Fast volumetric seizure model imaging in zebrafish brain with light sheet microscopy," Proc. SPIE 12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, 1284808 (12 March 2024); https://doi.org/10.1117/12.3000519