Fast Super-Resolution Imaging Technique and Immediate Early Nanostructure Capturing by a Photoconvertible Fluorescent Protein

Mingshu Zhang; Zhifei Fu; Changqing Li; Anyuan Liu; Dingming Peng; Fudong Xue; Wenting He; Shan Gao; Fan Xu; Dan Xu; Ling Yuan; Fa Zhang; Zhiheng Xu; Tao Xu; Pingyong Xu

doi:10.1021/acs.nanolett.9b02855

Fast Super-Resolution Imaging Technique and Immediate Early Nanostructure Capturing by a Photoconvertible Fluorescent Protein

Nano Lett. 2020 Apr 8;20(4):2197-2208. doi: 10.1021/acs.nanolett.9b02855. Epub 2019 Oct 7.

Authors

Mingshu Zhang¹, Zhifei Fu^{1

2}, Changqing Li¹, Anyuan Liu^{1

3}, Dingming Peng^{1

2}, Fudong Xue^{1

2}, Wenting He¹, Shan Gao^{2

4}, Fan Xu⁴, Dan Xu⁵, Ling Yuan⁶, Fa Zhang⁴, Zhiheng Xu^{2

7}, Tao Xu^{2

8}, Pingyong Xu^{1

2

8}

Affiliations

¹ Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
² College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100149, China.
³ School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, China.
⁴ Key Lab of Intelligent Information Processing, Institute of Computing Technology, Chinese Academy of Sciences, Beijing 100190, China.
⁵ College of Biological Science and Engineering, Institute of Life Sciences, Fuzhou University, Fuzhou 350116, China.
⁶ Center for Medical Genetics, School of Life Sciences, Central South University, Changsha, Hunan 410078, China.
⁷ State Key Laboratory of Molecular Developmental Biology, CAS Center for Excellence in Brain Science and Intelligence Technology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
⁸ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing100101, China.

PMID: 31576756
DOI: 10.1021/acs.nanolett.9b02855

Abstract

Low temporal resolution and limited photocontrollable fluorescent protein probes have restricted the widespread application of single-molecule localization microscopy (SMLM). In the current study, we developed a new photoconvertible fluorescent protein (PCFP), pcStar, and quick single molecule-guided Bayesian localization microscopy (Quick-SIMBA). The combination of pcStar and Quick-SIMBA achieved the highest temporal resolution (0.1-0.25 s) with large field-of-view (76 × 9.4 μm² -76 × 31.4 μm²) among the SMLM methods, which enabled the dynamic movements of the endoplasmic reticulum dense tubular matrix to be resolved. Moreover, pcStar extended the application of SMLM to imaging the immediate early nanostructures in Drosophila embryos and revealed a specific "parallel three-pillar" structure in the neuronal-glial cell junction, helping to elucidate glial cell "locking" and support of neurons during Drosophila embryogenesis.

Keywords: Drosophila embryos; Photoconvertible fluorescent protein; Quick-SIMBA; live super-resolution microscopy; pcStar.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actins / analysis
Animals
Bayes Theorem
Cell Line
Drosophila / embryology
Endoplasmic Reticulum / ultrastructure
Fluorescent Dyes / analysis*
Humans
Luminescent Proteins / analysis*
Microscopy, Fluorescence / methods
Single Molecule Imaging / methods*

Substances

Actins
Fluorescent Dyes
Luminescent Proteins