Article abstract
Nature Methods 5, 1047 - 1052 (2008)
Published online: 23 November 2008 | doi:10.1038/nmeth.1274
Whole-cell 3D STORM reveals interactions between cellular structures with nanometer-scale resolution
Bo Huang1,2,4, Sara A Jones2,4, Boerries Brandenburg1,2 & Xiaowei Zhuang1,2,3
Abstract
The ability to directly visualize nanoscopic cellular structures and their spatial relationship in all three dimensions will greatly enhance our understanding of molecular processes in cells. Here we demonstrated multicolor three-dimensional (3D) stochastic optical reconstruction microscopy (STORM) as a tool to quantitatively probe cellular structures and their interactions. To facilitate STORM imaging, we generated photoswitchable probes in several distinct colors by covalently linking a photoswitchable cyanine reporter and an activator molecule to assist bioconjugation. We performed 3D localization in conjunction with focal plane scanning and correction for refractive index mismatch to obtain whole-cell images with a spatial resolution of 20–30 nm and 60–70 nm in the lateral and axial dimensions, respectively. Using this approach, we imaged the entire mitochondrial network in fixed monkey kidney BS-C-1 cells, and studied the spatial relationship between mitochondria and microtubules. The 3D STORM images resolved mitochondrial morphologies as well as mitochondria-microtubule contacts that were obscured in conventional fluorescence images.
- Howard Hughes Medical Institute, Harvard University, 12 Oxford St. Cambridge, Massachusetts 02138, USA.
- Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St. Cambridge, Massachusetts 02138, USA.
- Department of Physics, Harvard University, 12 Oxford St. Cambridge, Massachusetts 02138, USA.
- These authors contributed equally to this work.
Correspondence to: Xiaowei Zhuang1,2,3 e-mail: zhuang@chemistry.harvard.edu
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