Ultra-High Resolution 3D Imaging of Whole Cells.

Huang, Fang; Sirinakis, George; Allgeyer, Edward S; Schroeder, Lena K; Duim, Whitney C; Kromann, Emil B; Phan, Thomy; Rivera-Molina, Felix E et al. · Cell · 2016

basic_science · Level V

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Abstract

Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. We demonstrate the wide applicability of W-4PiSMSN across diverse research fields by imaging complex molecular architectures ranging from bacteriophages to nuclear pores, cilia, and synaptonemal complexes in large 3D cellular volumes.

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