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Submitted on March 13, 2006
Accepted on August 2, 2006
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution
Eric Betzig 1*, George H. Patterson 2, Rachid Sougrat 2, O. Wolf Lindwasser 2, Scott Olenych 3, Juan S. Bonifacino 2, Michael W. Davidson 3, Jennifer Lippincott-Schwartz 2, Harald F. Hess 4
1 Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA; New Millennium Research LLC, Okemos, MI 48864, USA. 2 Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA. 3 National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310, USA. 4 NuQuest Research LLC, La Jolla, CA 92037, USA.
* To whom correspondence should be addressed.
Eric Betzig , E-mail: betzige{at}hhmi.org
A method for optically imaging intracellular proteins at nanometerspatial resolution is introduced. Numerous sparse subsets ofphotoactivatable fluorescent protein molecules are activated,localized (to ~2-25 nm), and then bleached. The aggregate positioninformation from all subsets is then assembled into a superresolutionimage. The method, termed photoactivated localization microscopy(PALM), is demonstrated in thin sections by imaging specifictarget proteins in lysosomes and mitochondria, and in fixed,whole cells by imaging vinculin at focal adhesions, actin withina lamellipodium, and the distribution of the retroviral proteinGag at the plasma membrane.
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