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Proceedings Paper

In vitro imaging of cells using peptide-conjugated quantum dots
Author(s): Mitsuru Ishikawa; Vasudevan Biju
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Paper Abstract

Efficient intracellular delivery of quantum dots (QDs) in living cells and elucidating the mechanism of the delivery are essential for advancing the applications of QDs to in vivo imaging and in vivo photodynamic therapy. Here, we demonstrate that clathrin-mediated endocytosis is the most dominant pathway for the delivery of peptide-conjugated QDs. We selected an insect neuropeptide, allatostatin (AST1), conjugated with CdSe-ZnS QDs, and investigated the delivery of the conjugate in living cells. We evaluated the contributions of clathrin-mediated endocytosis, receptormediated endocytosis, and charge-based cell penetration to the delivery of QD605-AST1 conjugates by flow cytometry and fluorescence video microscopy. The delivery was suppressed by ~57% in inhibiting phosphoinositide 3-kinase with wortmannin, which blocks the formation of clathrin-coated vesicles, and by ~45% in incubating the cells at 4°C. Also, we identified clathrin-mediated endocytosis by two-color experiment to find colocalization of QD560-labeled clathrin heavy-chain antibody and QD605-AST1. We further observed reduction of the galanin receptor-mediated delivery of QD605-AST1 by ~8% in blocking the cells with a galanin antagonist, and reduction of charge-based cell penetration delivery by ~30% in removing the positive charge in the peptide from arginine and suppressing the cell-surface negative charge from glycosaminoglycan.

Paper Details

Date Published: 17 February 2010
PDF: 8 pages
Proc. SPIE 7575, Colloidal Quantum Dots for Biomedical Applications V, 75750R (17 February 2010); doi: 10.1117/12.851055
Show Author Affiliations
Mitsuru Ishikawa, Health Technology Research Ctr., National Institute of Advanced Industrial Science and Technology (Japan)
Vasudevan Biju, Health Technology Research Ctr., National Institute of Advanced Industrial Science and Technology (Japan)

Published in SPIE Proceedings Vol. 7575:
Colloidal Quantum Dots for Biomedical Applications V
Marek Osinski; Wolfgang J. Parak; Thomas M. Jovin M.D.; Kenji Yamamoto M.D., Editor(s)

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