Eric Betzig1,2,3,4,5, George H. Patterson1,3,4,5, Rachid Sougrat1,3,4,5, O. Wolf Lindwasser1,3,4,5, Scott G. Olenych1,3,4,5, Juan S. Bonifacino1,3,4,5, Michael W. Davidson1,3,4,5, Jennifer Lippincott‐Schwartz1,3,4,5, Harald F. Hess1,2,3,4,5
1Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development (NICHD), Bethesda, MD 20892, USA.
2Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA
3National High Magnetic Field Laboratory, Florida State University, Tallahassee, Fl 32310 USA
4New Millennium Research, LLC, Okemos, MI 48864, USA.
5NuQuest Research, LLC, La Jolla, CA 92037, USA.
Tóm tắt
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 ∼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.
