Abstract
Super-resolution Radial Fluctuations (SRRF) imaging is a computational approach to fixed and live-cell super-resolution microscopy that is highly accessible to life science researchers since it uses common microscopes and open-source software plugins for ImageJ. This allows users to generate super-resolution images using the same equipment, fluorophores, fluorescent proteins and methods they routinely employ for their studies without specialized sample preparations or reagents. Here, we discuss a step-by-step workflow for acquiring and analyzing images using the NanoJ-SRRF software developed by the Ricardo Henriques group, with a focus on imaging chromatin. Increased accessibility of affordable super-resolution imaging techniques is an important step in extending the reach of this revolution in cellular imaging to a greater number of laboratories.
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Acknowledgments
Funding for microscope upgrades for SRRF imaging were funded by an Equipment Grant from the Dalhousie Medical Research Foundation (DMRF), and a Research, Tools, & Instruments (RTI) grant from the Natural Sciences and Engineering Research Council of Canada (NSERC). We would also like to thank Nvidia Corporation for the gift of the Titan V GPU used in this study obtained through their Higher Education and Research grants program. We would also like to thank Dr. Ricardo Henriques (Instituto Gulbenkian de Ciência, Portugal and University College London, UK) and his research group for their advice and access to the development-stage NanoJ-LiveSRRF software. Finally, we thank Dr. Gražvydas Lukinavičius and Dr. Jonas Bucevičius (Research group for chromatin labeling and imaging, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany) for the generous gift of the 5-TMR-Hoechst DNA dye used in this chapter.
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Salsman, J., Dellaire, G. (2022). Super-Resolution Radial Fluctuations (SRRF) Microscopy. In: Heit, B. (eds) Fluorescent Microscopy. Methods in Molecular Biology, vol 2440. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2051-9_14
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DOI: https://doi.org/10.1007/978-1-0716-2051-9_14
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