Dr. Huan-Cheng Chang - Correlative Light-Electron Microscopy of Lipid-Encapsulated Fluorescent Nan

Correlative Light-Electron Microscopy of Lipid-Encapsulated Fluorescent Nanodiamonds for Nanometric Localization of Cell Surface Antigens

Feng-Jen Hsieh,^1-3,§ Yen-Wei Chen,^1,§ Yao-Kuan Huang,⁴ Hsien-Ming Lee,⁵ Chun-Hung Lin,^2,3,6 and Huan-Cheng Chang*^,1,7

¹Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan

²Taiwan International Graduate Program – Chemical Biology and Molecular Biophysics, Academia Sinica, Taipei 115, Taiwan

³Department of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan

⁴Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan

⁵Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan

⁶Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan

⁷Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan

^§These two authors contribute equally to this work.

Anal. Chem. 90, 1566–1571 (2018).

Containing 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 sub-diffraction 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 ~50 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. [link]

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