Simultaneous multicolor detection system of the single-molecular microbial antigen with total internal reflection fluorescence microscopy.
Microbiol Immunol
; 49(5): 461-70, 2005.
Article
en En
| MEDLINE
| ID: mdl-15905608
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 nanocrystal 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.
Buscar en Google
Bases de datos:
MEDLINE
Asunto principal:
Coloración y Etiquetado
/
Toxina Tetánica
/
Técnica del Anticuerpo Fluorescente
/
Toxina Diftérica
/
Microscopía Fluorescente
/
Antígenos
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Microbiol Immunol
Año:
2005
Tipo del documento:
Article