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Science. 2006 Sep 15;313(5793):1642-5. Epub 2006 Aug 10.

Imaging intracellular fluorescent proteins at nanometer resolution.

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  • 1Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org

Abstract

We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.

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  • The limits of light. [Nat Rev Mol Cell Biol. 2010]
PMID:
16902090
[PubMed - indexed for MEDLINE]
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