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

Xu, Ke; Babcock, Hazen P; Zhuang, Xiaowei · Nat Methods · 2012

basic_science · Level V

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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.

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