Fast Super-Resolution Imaging Technique and Immediate Early Nanostructure Capturing by a Photoconvertible Fluorescent Protein.
Mingshu ZhangZhifei FuChangqing LiAnyuan LiuDingming PengFudong XueWenting HeShan GaoFan XuDan XuLing YuanFa ZhangZhiheng XuTao XuPingyong XuPublished in: Nano letters (2019)
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.
Keyphrases
- single molecule
- living cells
- single cell
- atomic force microscopy
- endoplasmic reticulum
- quantum dots
- high resolution
- protein protein
- cell therapy
- spinal cord
- binding protein
- neuropathic pain
- stem cells
- small molecule
- label free
- fluorescence imaging
- spinal cord injury
- endothelial cells
- mesenchymal stem cells
- high glucose