Super-Resolution Radial Fluctuations (SRRF) Microscopy

Methods Mol Biol. 2022;2440:225-251. doi: 10.1007/978-1-0716-2051-9_14.

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.

Keywords: Chromatin; EMCCD; SRRF; Super-resolution; sCMOS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Dyes*
  • Ionophores
  • Microscopy, Fluorescence / methods
  • Software*

Substances

  • Fluorescent Dyes
  • Ionophores