Unit

UNIT 21.8 Superresolution Imaging with Standard Fluorescent Probes

  1. Bryan A. Millis1,
  2. Dylan T. Burnette2,
  3. Jennifer Lippincott-Schwartz2,
  4. Bechara Kachar1

Published Online: 24 SEP 2013

DOI: 10.1002/0471143030.cb2108s60

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Millis, B. A., Burnette, D. T., Lippincott-Schwartz, J. and Kachar, B. 2013. Superresolution Imaging with Standard Fluorescent Probes. Current Protocols in Cell Biology. 60:21.8:21.8.1–21.8.17.

Author Information

  1. 1

    Laboratory of Cell Structure and Dynamics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland

  2. 2

    The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 24 SEP 2013

Abstract

For more than 100 years, the ultimate resolution of a light microscope (∼200 nm) has been constrained by the fundamental physical phenomenon of diffraction, as described by Ernst Abbe in 1873. While this limitation is just as applicable to today's light microscopes, it is the combination of high-end optics, clever methods of sample illumination, and computational techniques that has enabled researchers to access information at an order of magnitude greater resolution than once thought possible. This combination, broadly termed superresolution microscopy, has been increasingly practical for many labs to implement from both a hardware and software standpoint, but, as with many cutting-edge techniques, it also comes with limitations. One of the current drawbacks to superresolution microscopy is the limited number of probes and conditions that have been suitable for imaging. Here, a technique termed bleaching/blinking-assisted localization microscopy (BaLM) makes use of the inherent blinking and bleaching properties of almost all fluorophores as a means to generate superresolution images. Curr. Protoc. Cell Biol. 60:21.8.1-21.8.17. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • superresolution;
  • BaLM;
  • microscopy;
  • bleaching;
  • blinking