| Autor: |
Yang Zhang, Gaoxiang Wang, Peizhou Huang, Edison Sun, Junghun Kweon, Qianru Li, Ji Zhe, Leslie L. Ying, Hao F. Zhang |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Anal Chem |
| ISSN: |
1520-6882 |
| Popis: |
Super-resolution microscopy can capture spatiotemporal organizations of protein interactions with resolution down to 10 nm; however, the analyses of more than two proteins involving low-abundance protein are challenging because spectral crosstalk and heterogeneities of individual fluorescent labels result in molecular misidentification. Here we developed a deep learning-based imaging analysis method for spectroscopic single-molecule localization microscopy to minimize molecular misidentification in three-color super-resolution imaging. We characterized the 3-fold reduction of molecular misidentification in the new imaging method using pure samples of different photoswitchable fluorophores and visualized three distinct subcellular proteins in U2-OS cell lines. We further validated the protein counts and interactions of TOMM20, DRP1, and SUMO1 in a well-studied biological process - Staurosporine-induced apoptosis by comparing the imaging results with Western-blot analysis of different subcellular portions. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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