Intracellular imaging of targeted proteins labeled with quantum dots

We developed a new method for imaging the movement of targeted proteins in living cancer cells with photostable and bright quantum dots (QDs). QDs were conjugated with various molecules and proteins, such as phalloidin, anti-tubulin antibody and kinesin. These bioconjugated QDs were mixed with a tra...

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Bibliographic Details
Published in:Experimental cell research 2008-11, Vol.314 (19), p.3563-3569
Main Authors: Yoo, Jungwoo, Kambara, Taketoshi, Gonda, Kohsuke, Higuchi, Hideo
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
ACTIN
ANTIBODIES
CARCINOMAS
Cellular biology
Imaging
Imaging, Three-Dimensional - methods
IN VITRO
Kinesin
Kinesin - metabolism
Microscope
Microscopy, Confocal
Microscopy, Fluorescence
Microtubule
MICROTUBULES
Motor protein
Phalloidine - metabolism
Protein Transport
Proteins
Proteins - analysis
Proteins - metabolism
QUANTUM DOTS
Research methodology
Scientific imaging
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Summary:We developed a new method for imaging the movement of targeted proteins in living cancer cells with photostable and bright quantum dots (QDs). QDs were conjugated with various molecules and proteins, such as phalloidin, anti-tubulin antibody and kinesin. These bioconjugated QDs were mixed with a transfection reagent and successfully internalized into living cells. The movements of individual QDs were tracked for long periods of time. Phalloidin conjugated QDs bound to actin filaments and showed almost no movement. In contrast, anti-tubulin antibody conjugated QDs bound to microtubules and revealed dynamic movement of microtubules. Kinesin showed an interesting behavior whereby kinesin came to be almost paused briefly for a few seconds and then moved once again. This is in direct contrast to the smoothly continuous movement of kinesin in an in vitro assay. The maximum velocity of kinesin in cells was faster than that in the in vitro assay. These results suggest that intracellular movement of kinesin is different from that in the in vitro assay. This newly described method will be a powerful tool for investigating the functions of proteins in living cells.
ISSN:0014-4827
1090-2422
DOI:10.1016/j.yexcr.2008.09.014