Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 967-983 |
| Number of pages | 17 |
| Journal | Journal of Cell Biology |
| Volume | 202 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2013 |
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ASJC Scopus subject areas
- Cell Biology
Cite this
Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging. / Nishimura, Hirohito; Ritchie, Ken; Kasai, Rinshi S.; Goto, Miki; Morone, Nobuhiro; Sugimura, Hiroyuki; Tanaka, Koichiro; Sase, Ichiro; Yoshimura, Akihiko; Nakano, Yoshitaro; Fujiwara, Takahiro K.; Kusumi, Akihiro.
In: Journal of Cell Biology, Vol. 202, No. 6, 2013, p. 967-983.Research output: Contribution to journal › Article
}
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.
UR - http://www.scopus.com/inward/record.url?scp=84884376017&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84884376017&partnerID=8YFLogxK
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 -