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Document Version Version Comment Date Status
732422.0 Automatic journal name synchronization 29.04.2017 20:16 Released

ID: 732422.0, MPI für molekulare Zellbiologie und Genetik / MPI-CBG Publications 2016 (archival)
Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development
Authors:Icha, Jaroslav; Schmied, Christopher; Sidhaye, Jaydeep; Tomancák, Pavel; Preibisch, Stephan; Norden, Caren
Date of Publication (YYYY-MM-DD):2016
Title of Journal:Journal of Visualized Experiments : Jove
Issue / Number:110
Sequence Number of Article:e53966
Copyright:not available
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:Light sheet fluorescence microscopy (LSFM) is gaining more and more popularity as a method to image embryonic development. The main advantages of LSFM compared to confocal systems are its low phototoxicity, gentle mounting strategies, fast acquisition with high signal to noise ratio and the possibility of imaging samples from various angles (views) for long periods of time. Imaging from multiple views unleashes the full potential of LSFM, but at the same time it can create terabyte-sized datasets. Processing such datasets is the biggest challenge of using LSFM. In this protocol we outline some solutions to this problem. Until recently, LSFM was mostly performed in laboratories that had the expertise to build and operate their own light sheet microscopes. However, in the last three years several commercial implementations of LSFM became available, which are multipurpose and easy to use for any developmental biologist. This article is primarily directed to those researchers, who are not LSFM technology developers, but want to employ LSFM as a tool to answer specific developmental biology questions. Here, we use imaging of zebrafish eye development as an example to introduce the reader to LSFM technology and we demonstrate applications of LSFM across multiple spatial and temporal scales. This article describes a complete experimental protocol starting with the mounting of zebrafish embryos for LSFM. We then outline the options for imaging using the commercially available light sheet microscope. Importantly, we also explain a pipeline for subsequent registration and fusion of multiview datasets using an open source solution implemented as a Fiji plugin. While this protocol focuses on imaging the developing zebrafish eye and processing data from a particular imaging setup, most of the insights and troubleshooting suggestions presented here are of general use and the protocol can be adapted to a variety of light sheet microscopy experiments.
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Thüm
Affiliations:MPI für molekulare Zellbiologie und Genetik