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Updated: Jun 1, 2025
Author Spotlight: In Vivo Whole-Brain Imaging of Zebrafish Larvae Using Three-Dimensional Fluorescence Microscopy
Published on: April 28, 2023
Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount
Olga Doszyn1, Tomasz Dulski2, Justyna Zmorzynska1
1Laboratory of Developmental Neurobiology, International Institute of Molecular Mechanisms and Machines, 02-247 Warsaw, Poland; Laboratory of Molecular and Cellular Neurobiology, International Institute of Molecular and Cell Biology in Warsaw, 02-109 Warsaw, Poland.
Abstract:
Due to their small size and transparency, larval zebrafish are a useful model for whole-brain imaging. Here, we present a protocol for the visualization of phosphorylated Rps6, a marker of mechanistic target of rapamycin complex 1 (mTORC1) activity, in the zebrafish brains at 5 days post fertilization (dpf), using whole-mount immunofluorescence and light-sheet microscopy. We describe steps for sample preparation, storage, staining, and imaging. This protocol can also be modified for staining with antibodies against other proteins. For complete details on the use and execution of this protocol, please refer to Doszyn et al.1.
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