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Mol Biol Cell. 2018 Jul 1;29(13):1519-1525. doi: 10.1091/mbc.E17-05-0276.

A beginner's guide to rigor and reproducibility in fluorescence imaging experiments.

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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.


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.

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