ADVANCED IMAGING. Extended-resolution Structured Illumination Imaging of Endocytic and Cytoskeletal Dynamics

Overview

Journal Science

Specialty Science

Date 2015 Aug 29

PMID 26315442

Citations 267

Authors

Dong Li

Lin Shao

Bi-Chang Chen

Xi Zhang

Mingshu Zhang

Brian Moses

Daniel E Milkie

Jordan R Beach

John A Hammer 3rd

Mithun Pasham

Tomas Kirchhausen

Michelle A Baird

Michael W Davidson

Pingyong Xu

Eric Betzig

Affiliations

Soon will be listed here.

Abstract

Super-resolution fluorescence microscopy is distinct among nanoscale imaging tools in its ability to image protein dynamics in living cells. Structured illumination microscopy (SIM) stands out in this regard because of its high speed and low illumination intensities, but typically offers only a twofold resolution gain. We extended the resolution of live-cell SIM through two approaches: ultrahigh numerical aperture SIM at 84-nanometer lateral resolution for more than 100 multicolor frames, and nonlinear SIM with patterned activation at 45- to 62-nanometer resolution for approximately 20 to 40 frames. We applied these approaches to image dynamics near the plasma membrane of spatially resolved assemblies of clathrin and caveolin, Rab5a in early endosomes, and α-actinin, often in relationship to cortical actin. In addition, we examined mitochondria, actin, and the Golgi apparatus dynamics in three dimensions.

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