Nuclear pores as versatile reference standards for quantitative superresolution microscopy

Authors: Jervis Vermal Thevathasan, Maurice Kahnwald, Konstanty Cieśliński, Philipp Hoess, Sudheer Kumar Peneti, Manuel Reitberger, Daniel Heid, Krishna Chaitanya Kasuba, Sarah Janice Hoerner, Yiming Li, Yu-Le Wu, Markus Mund, Ulf Matti, Pedro Matos Pereira, Ricardo Henriques, Bianca Nijmeijer, Moritz Kueblbeck, Vilma Jimenez Sabinina, Jan Ellenberg, Jonas Ries
Technologies: CARE, NanoJ, NanoJ-SQUIRREL, NanoJ-SRRF and Nuclear-Pores as references
Paper published in Nature Methods, September 2019
Publisher: Nature Publishing Group US New York

The manuscript describes various studies focusing on the use of nuclear pore complexes (NPCs) as reference structures for quantitative superresolution microscopy. Researchers generated stable cell lines in human and yeast cells, where specific nucleoporins were endogenously tagged with different fluorescent labels. They validated the approach by counting protein copy numbers and addressing limitations. The studies aim to provide valuable resources for the community to assess microscope performance, optimize imaging conditions, and determine effective labeling efficiencies. The manuscript also includes methods for genome editing, expansion microscopy, and nanobody labeling, as well as data analysis using custom code and various software. The data will be accessible on BioStudies, and the antibodies and cell lines used in the study were properly characterized.