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Nat Methods. 2012 Jan 8;9(2):185-8. doi: 10.1038/nmeth.1841.

Dual-objective STORM reveals three-dimensional filament organization in the actin cytoskeleton.

Author information

1
Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.

Abstract

By combining astigmatism imaging with a dual-objective scheme, we improved the image resolution of stochastic optical reconstruction microscopy (STORM) and obtained <10-nm lateral resolution and <20-nm axial resolution when imaging biological specimens. Using this approach, we resolved individual actin filaments in cells and revealed three-dimensional ultrastructure of the actin cytoskeleton. We observed two vertically separated layers of actin networks with distinct structural organizations in sheet-like cell protrusions.

PMID:
22231642
PMCID:
PMC3304438
DOI:
10.1038/nmeth.1841
[Indexed for MEDLINE]
Free PMC Article

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