Correlative Light-Electron Microscopy of Lipid-Encapsulated Fluorescent Nanodiamonds for Nanometric Localization of Cell Surface Antigens
At a Glance
Section titled âAt a Glanceâ| Metadata | Details |
|---|---|
| Publication Date | 2017-12-19 |
| Journal | Analytical Chemistry |
| Authors | FengâJen Hsieh, YenâWei Chen, Yao-Kuan Huang, HsienâMing Lee, ChunâHung Lin |
| Institutions | Institute of Cellular and Organismic Biology, Academia Sinica, Institute of Biological Chemistry, Academia Sinica |
| Citations | 35 |
Abstract
Section titled âAbstractâContaining an ensemble of nitrogen-vacancy centers in crystal matrices, fluorescent nanodiamonds (FNDs) are a new type of photostable markers that have found wide applications in light microscopy. The nanomaterial also has a dense carbon core, making it visible to electron microscopy. Here, we show that FNDs encapsulated in biotinylated lipids (bLs) are useful for subdiffraction imaging of antigens on cell surface with correlative light-electron microscopy (CLEM). The lipid encapsulation enables not only good dispersion of the particles in biological buffers but also high specific labeling of live cells. By employing the bL-encapsulated FNDs to target CD44 on HeLa cell surface through biotin-mediated immunostaining, we obtained the spatial distribution of these antigens by CLEM with a localization accuracy of âź50 nm in routine operations. A comparative study with dual-color imaging, in which CD44 was labeled with FND and MICA/MICB was labeled with Alexa Fluor 488, demonstrated the superior performance of FNDs as fluorescent fiducial markers for CLEM of cell surface antigens.
Tech Support
Section titled âTech SupportâOriginal Source
Section titled âOriginal SourceâReferences
Section titled âReferencesâ- 2012 - Correlative Light and Electron Microscopy