A starter kit for point-localization super-resolution imaging

Super-resolution fluorescence imaging can be achieved through the localization of single molecules. By using suitable dyes, optical configurations, and software, it is possible to study a wide variety of biological systems. Here, we summarize the different approaches to labeling proteins. We review proven imaging modalities, and the features of freely available software. Finally, we give an overview of some biological applications. We conclude by synthesizing these different technical aspects into recommendations for standards that the field might apply to ensure quality of images and comparability of algorithms and dyes.

► Super-resolution is possible with fluorescent protein labels and many synthetic dyes.
► Freely available and open source software can be used to analyze data.
► Imaging modalities include 3D, live, and multicolor imaging.
► Super-resolution has been applied to answer diverse biological questions.