| Autor: |
Hsieh FJ; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan.; Institute of Biochemical Sciences , National Taiwan University , Taipei 106 , Taiwan., Sotoma S; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan., Lin HH; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan., Cheng CY; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan., Yu TY; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan., Hsieh CL; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan., Lin CH; Institute of Biochemical Sciences , National Taiwan University , Taipei 106 , Taiwan., Chang HC; Institute of Atomic and Molecular Sciences , Academia Sinica , Taipei 106 , Taiwan.; Department of Chemical Engineering , National Taiwan University of Science and Technology , Taipei 106 , Taiwan.; Department of Chemistry , National Taiwan Normal University , Taipei 106 , Taiwan. |
| Abstrakt: |
Real-time tracking of membrane proteins is essential to gain an in-depth understanding of their dynamics on the cell surface. However, conventional fluorescence imaging with molecular probes like organic dyes and fluorescent proteins often suffers from photobleaching of the fluorophores, thus hindering their use for continuous long-term observations. With the availability of fluorescent nanodiamonds (FNDs), which have superb biocompatibility and excellent photostability, it is now possible to conduct the imaging in both short and long terms with high temporal and spatial resolution. To realize the concept, we have developed a facile method (e.g., one-pot preparation) to produce alkyne-functionalized hyperbranched-polyglycerol-coated FNDs for bioorthogonal labeling of azide-modified membrane proteins and azide-modified antibodies of membrane proteins. The high specificity of this labeling method has allowed us to continuously monitor the movements of the proteins of interest (such as integrin α5) on/in living cells over 2 h. The results open a new horizon for live cell imaging with functional nanoparticles and fluorescence microscopy. |
