Journal
MICRON
Volume 41, Issue 3, Pages 198-202Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.micron.2009.11.002
Keywords
Near-field scanning microscopy (NSOM); Quantum dot (QD); Mesenchymal stem cells (MSCs); CD44
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Funding
- National Natural Science Foundation of China [30872404]
- Sino-America joint [30828028]
- National Institute of Health [R01 HL60635]
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The lateral organization of membrane proteins and lipids domains has a direct impact on many cellular processes, but generally these domains are too small to be resolved by diffraction-limited resolution of fluorescence microscopy. Here, we use quantum dot (QD) labeling based on near-field optical imaging, to study the nanoscale organization of hyaluronan receptor CD44 molecules of fixed mesenchymal stem cells (MSCs) in air, with a optical resolution down to 50 nm. The photostability and high luminance of QD evidently improve the signal-to-noise ratio and reproducibility of near-field optical data. Importantly, the blinking-intensity analysis was proposed to identify single QD, providing a calibration to relate intensity to numbers of antibody for the first time. Additionally, the fluorescence-topographic imaging enables us to investigate the topographic location pattern. Our results demonstrate that CD44 molecules on MSCs are enriched into nanosized domain and they predominantly locate on the peak of the membrane protrusions, which may contribute to clarify the underlying mechanism of functions ascribed to these molecules. (C) 2009 Elsevier Ltd. All rights reserved.
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