Format

Send to

Choose Destination
See comment in PubMed Commons below
Cell. 2016 Aug 11;166(4):1028-1040. doi: 10.1016/j.cell.2016.06.016. Epub 2016 Jul 7.

Ultra-High Resolution 3D Imaging of Whole Cells.

Author information

1
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA.
2
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; The Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
3
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
4
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Chemistry, Harvey Mudd College, Claremont, CA 91711, USA.
5
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Biomedical Engineering, Yale University, CT 06520, USA.
6
Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
7
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
8
Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA; Howard Hughes Medical Institute, Yale University, New Haven, CT 06520, USA; Department of Microbial Pathogenesis, Yale School of Medicine, New Haven, CT 06520, USA.
9
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Nanobiology Institute, Yale University, West Haven, CT 06516, USA.
10
Department of Engineering Science, University of Oxford, Oxford OX1 3PJ, UK; Centre for Neural Circuits and Behaviour, University of Oxford, Oxford OX1 3SR, UK.
11
Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Biomedical Engineering, Yale University, CT 06520, USA; Nanobiology Institute, Yale University, West Haven, CT 06516, USA. Electronic address: joerg.bewersdorf@yale.edu.

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.

PMID:
27397506
PMCID:
PMC5005454
DOI:
10.1016/j.cell.2016.06.016
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Support Center