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Proceedings Paper

Simultaneous multicolor detection system of the single-molecular microbial antigen by total internal reflection fluorescence microscopy with fluorescent nanocrystal quantum dots
Author(s): Akiyoshi Hoshino; Kouki Fujioka; Mayu Yamamoto; Noriyoshi Manabe; Masato Yasuhara; Kazuo Suzuki; Kenji Yamamoto
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Paper Abstract

Immunological diagnostic methods have been widely performed and showed high performance in molecular and cellular biology, molecular imaging, and medical diagnostics. We have developed novel methods for the fluorescent labeling of several antibodies coupled with fluorescent nanocrystals QDs. In this study we demonstrated that two bacterial toxins, diphtheria toxin and tetanus toxin, were detected simultaneously in the same view field of a cover slip by using directly QD-conjugated antibodies. We have succeeded in detecting bacterial toxins by counting luminescent spots on the evanescent field with using primary antibody conjugated to QDs. In addition, each bacterial toxin in the mixture can be separately detected by single excitation laser with emission band pass filters, and simultaneously in situ pathogen quantification was performed by calculating the luminescent density on the surface of the cover slip. Our results demonstrate that total internal reflection fluorescence microscopy (TIRFM) enables us to distinguish each antigen from mixed samples and can simultaneously quantitate multiple antigens by QD-conjugated antibodies. Bioconjugated QDs could have great potentialities for in practical biomedical applications to develop various high-sensitivity detection systems.

Paper Details

Date Published: 21 November 2005
PDF: 11 pages
Proc. SPIE 6009, Optical Methods in Drug Discovery and Development, 60090P (21 November 2005); doi: 10.1117/12.630344
Show Author Affiliations
Akiyoshi Hoshino, International Medical Ctr. of Japan (Japan)
Tokyo Medical and Dental Univ. (Japan)
Kouki Fujioka, International Medical Ctr. of Japan (Japan)
Mayu Yamamoto, International Medical Ctr. of Japan (Japan)
Noriyoshi Manabe, International Medical Ctr. of Japan (Japan)
Masato Yasuhara, Tokyo Medical and Dental Univ. (Japan)
Kazuo Suzuki, National Institute of Infectious Diseases (Japan)
Kenji Yamamoto, International Medical Ctr. of Japan (Japan)
Tokyo Medical and Dental Univ. (Japan)


Published in SPIE Proceedings Vol. 6009:
Optical Methods in Drug Discovery and Development
Mostafa Analoui; David A. Dunn, Editor(s)

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