TY - JOUR
T1 - Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging
AU - Nishimura, Hirohito
AU - Ritchie, Ken
AU - Kasai, Rinshi S.
AU - Goto, Miki
AU - Morone, Nobuhiro
AU - Sugimura, Hiroyuki
AU - Tanaka, Koichiro
AU - Sase, Ichiro
AU - Yoshimura, Akihiko
AU - Nakano, Yoshitaro
AU - Fujiwara, Takahiro K.
AU - Kusumi, Akihiro
PY - 2013
Y1 - 2013
N2 - Fluorescence microscopy is used extensively in cellbiological and biomedical research, but it is often plagued by three major problems with the presently available fluorescent probes: photobleaching, blinking, and large size. We have addressed these problems, with special attention to single-molecule imaging, by developing biocompatible, red-emitting silicon nanocrystals (SiNCs) with a 4.1-nm hydrodynamic diameter. Methods for producing SiNCs by simple chemical etching, for hydrophilically coating them, and for conjugating them to biomolecules precisely at a 1:1 ratio have been developed. Single SiNCs neither blinked nor photobleached during a 300-min overall period observed at video rate. Single receptor molecules in the plasma membrane of living cells (using transferrin receptor) were imaged for =10 times longer than with other probes, making it possible for the first time to observe the internalization process of receptor molecules at the single-molecule level. Spatial variations of molecular diffusivity in the scale of 1-2 μm, i.e., a higher level of domain mosaicism in the plasma membrane, were revealed.
AB - Fluorescence microscopy is used extensively in cellbiological and biomedical research, but it is often plagued by three major problems with the presently available fluorescent probes: photobleaching, blinking, and large size. We have addressed these problems, with special attention to single-molecule imaging, by developing biocompatible, red-emitting silicon nanocrystals (SiNCs) with a 4.1-nm hydrodynamic diameter. Methods for producing SiNCs by simple chemical etching, for hydrophilically coating them, and for conjugating them to biomolecules precisely at a 1:1 ratio have been developed. Single SiNCs neither blinked nor photobleached during a 300-min overall period observed at video rate. Single receptor molecules in the plasma membrane of living cells (using transferrin receptor) were imaged for =10 times longer than with other probes, making it possible for the first time to observe the internalization process of receptor molecules at the single-molecule level. Spatial variations of molecular diffusivity in the scale of 1-2 μm, i.e., a higher level of domain mosaicism in the plasma membrane, were revealed.
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U2 - 10.1083/jcb.201301053
DO - 10.1083/jcb.201301053
M3 - Article
C2 - 24043702
AN - SCOPUS:84884376017
VL - 202
SP - 967
EP - 983
JO - Journal of Cell Biology
JF - Journal of Cell Biology
SN - 0021-9525
IS - 6
ER -