Molecular Imaging Center, Cancer Research Laboratory, University of California, Berkeley, Berkeley, CA 94720.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720.
Mol Biol Cell. 2018 Jul 1;29(13):1519-1525. doi: 10.1091/mbc.E17-05-0276.
Fluorescence light microscopy is an indispensable approach for the investigation of cell biological mechanisms. With the development of cutting-edge tools such as genetically encoded fluorescent proteins and superresolution methods, light microscopy is more powerful than ever at providing insight into a broad range of phenomena, from bacterial fission to cancer metastasis. However, as with all experimental approaches, care must be taken to ensure reliable and reproducible data collection, analysis, and reporting. Each step of every imaging experiment, from design to execution to communication to data management, should be critically assessed for bias, rigor, and reproducibility. This Perspective provides a basic "best practices" guide for designing and executing fluorescence imaging experiments, with the goal of introducing researchers to concepts that will help empower them to acquire images with rigor.
荧光显微镜是研究细胞生物学机制不可或缺的方法。随着基因编码荧光蛋白和超分辨率方法等尖端工具的发展，荧光显微镜在提供对各种现象的深入了解方面变得比以往任何时候都更加强大，从细菌分裂到癌症转移。然而，与所有实验方法一样，必须小心谨慎，以确保可靠和可重复的数据收集、分析和报告。从设计到执行，从交流到数据管理，每个成像实验的步骤都必须严格评估其偏差、严谨性和可重复性。本观点提供了设计和执行荧光成像实验的基本“最佳实践”指南，旨在向研究人员介绍有助于他们严格获取图像的概念。
