Label-retention expansion microscopy
| Autor: | Bo Huang, Zhipeng Dai, Xiaoyu Shi, Xiaomeng Wang, Dhivya Ashok Kumar, Tracy T. Chow, Alejandro Ramirez, Jeremy F. Reiter, Eric J. Huang, Andrew McColloch, Qi Li, Siyu Feng, Zixi Lin, Ian B. Seiple, Arthur A Tran, Jiapei Chen |
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| Rok vydání: | 2021 |
| Předmět: |
In situ
Microscope Biotin Succinimides Biology Molecular resolution Microtubules Signal Antibodies law.invention Mice 03 medical and health sciences 0302 clinical medicine law Cell Line Tumor Microscopy Animals Humans Fluorescent Dyes 030304 developmental biology 0303 health sciences Osteoblasts Staining and Labeling Mouse Embryonic Stem Cells Cell Biology Superresolution Fluorescence Optical reconstruction HEK293 Cells Microscopy Fluorescence Streptavidin 030217 neurology & neurosurgery HeLa Cells Biomedical engineering |
| Zdroj: | Journal of Cell Biology. 220 |
| ISSN: | 1540-8140 0021-9525 |
| DOI: | 10.1083/jcb.202105067 |
| Popis: | Expansion microscopy (ExM) increases the effective resolving power of any microscope by expanding the sample with swellable hydrogel. Since its invention, ExM has been successfully applied to a wide range of cell, tissue, and animal samples. Still, fluorescence signal loss during polymerization and digestion limits molecular-scale imaging using ExM. Here, we report the development of label-retention ExM (LR-ExM) with a set of trifunctional anchors that not only prevent signal loss but also enable high-efficiency labeling using SNAP and CLIP tags. We have demonstrated multicolor LR-ExM for a variety of subcellular structures. Combining LR-ExM with superresolution stochastic optical reconstruction microscopy (STORM), we have achieved molecular resolution in the visualization of polyhedral lattice of clathrin-coated pits in situ. |
| Databáze: | OpenAIRE |
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