Imaging Intracellular Fluorescent Proteins at Nanometer Resolution
Florida State University · Howard Hughes Medical Institute · +5 more institutions
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.
Citation impact
- FWCI
- 210.87
- Percentile
- 100%
- References
- 29
Authors
9- EBEric BetzigCorresponding
Florida State University, Howard Hughes Medical Institute, Janelia Research Campus, ProQuest (United States), National High Magnetic Field Laboratory, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Millennium Engineering and Integration (United States)
- GHGeorge H. Patterson
Florida State University, Howard Hughes Medical Institute, Janelia Research Campus, ProQuest (United States), National High Magnetic Field Laboratory, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Millennium Engineering and Integration (United States)
- RSRachid Sougrat
Florida State University, Howard Hughes Medical Institute, Janelia Research Campus, ProQuest (United States), National High Magnetic Field Laboratory, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Millennium Engineering and Integration (United States)
- OWO. Wolf Lindwasser
Florida State University, Howard Hughes Medical Institute, Janelia Research Campus, ProQuest (United States), National High Magnetic Field Laboratory, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Millennium Engineering and Integration (United States)
- SGScott G. Olenych
Florida State University, Howard Hughes Medical Institute, Janelia Research Campus, ProQuest (United States), National High Magnetic Field Laboratory, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Millennium Engineering and Integration (United States)
Topics & keywords
- Vinculin
- Lamellipodium
- Microscopy
- Actin
- Superresolution
- Resolution (logic)
- Super-resolution microscopy
- Photoactivated localization microscopy