Single-shot super-resolution total internal reflection fluorescence microscopy

Min Guo; Panagiotis Chandris; John Paul Giannini; Adam J Trexler; Robert Fischer; Jiji Chen; Harshad D Vishwasrao; Ivan Rey-Suarez; Yicong Wu; Xufeng Wu; Clare M Waterman; George H Patterson; Arpita Upadhyaya; Justin W Taraska; Hari Shroff

doi:10.1038/s41592-018-0004-4

Single-shot super-resolution total internal reflection fluorescence microscopy

Nat Methods. 2018 Jun;15(6):425-428. doi: 10.1038/s41592-018-0004-4. Epub 2018 May 7.

Authors

Min Guo¹, Panagiotis Chandris², John Paul Giannini^{2

3}, Adam J Trexler^{4

5}, Robert Fischer⁴, Jiji Chen⁶, Harshad D Vishwasrao⁶, Ivan Rey-Suarez^{2

3}, Yicong Wu², Xufeng Wu⁴, Clare M Waterman⁴, George H Patterson⁷, Arpita Upadhyaya⁸, Justin W Taraska⁴, Hari Shroff^{2

6

8}

Affiliations

¹ Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA. min.guo@nih.gov.
² Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.
³ Biophysics Program, University of Maryland, College Park, MD, USA.
⁴ National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
⁵ Northrop Grumman Corporation, Monterey, CA, USA.
⁶ Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, MD, USA.
⁷ Section on Biophotonics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.
⁸ Department of Physics and Institute for Physical Science and Technology, University of Maryland, College Park, MD, USA.

Abstract

We combined instant structured illumination microscopy (iSIM) with total internal reflection fluorescence microscopy (TIRFM) in an approach referred to as instant TIRF-SIM, thereby improving the lateral spatial resolution of TIRFM to 115 ± 13 nm without compromising speed, and enabling imaging frame rates up to 100 Hz over hundreds of time points. We applied instant TIRF-SIM to multiple live samples and achieved rapid, high-contrast super-resolution imaging close to the coverslip surface.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Line, Tumor
Humans
Microscopy, Fluorescence / methods*
Microtubules
Osteosarcoma
rab GTP-Binding Proteins / physiology

Substances

rab11 protein
rab GTP-Binding Proteins

Grants and funding

Z99 EB999999/ImNIH/Intramural NIH HHS/United States