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Simple detection of small amounts of Pseudomonas cells in milk by using a microfluidic device.
Yamaguchi, N; Ohba, H; Nasu, M.
Afiliación
  • Yamaguchi N; Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
Lett Appl Microbiol ; 43(6): 631-6, 2006 Dec.
Article en En | MEDLINE | ID: mdl-17083709
AIMS: Flow cytometry offers rapid and reliable analyses of bacteria in milk. However, a flow cytometer is relatively expensive and operation is rather complicated for an unskilled operator. We applied flow cytometry using a microfluidic device (on-chip flow cytometry) in detection of small amounts of milk-spoiling bacteria. METHODS AND RESULTS: Pseudomonas cells in milk were in situ hybridized with Cy5-labelled probe specific for Pseudomonas spp. under optimized condition. Numbers of Pseudomonas cells in the stationary phase and in the starved state determined by on-chip flow cytometry were compared with those determined by conventional plate counting, and on-chip flow cytometry detected targeted cells in milk that were undetectable as colony forming units(CFU) on Standards Methods Agar. CONCLUSIONS: The contamination in milk with fewer than 10 CFU ml(-1) of targeted cells in starved state was detectable with simple procedure (0.5 h milk-clearing, 1 h fixation, 2 h hybridization and 0.5 h on-chip flow cytometry following 12 h enrichment of cells). SIGNIFICANCE AND IMPACT OF THE STUDY: On-chip flow cytometry following fluorescence in situ hybridization could be applicable to simple detection of milk-spoiling bacteria.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pseudomonas / Hibridación Fluorescente in Situ / Leche / Técnicas Analíticas Microfluídicas / Citometría de Flujo / Microbiología de Alimentos Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: Lett Appl Microbiol Asunto de la revista: MICROBIOLOGIA Año: 2006 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pseudomonas / Hibridación Fluorescente in Situ / Leche / Técnicas Analíticas Microfluídicas / Citometría de Flujo / Microbiología de Alimentos Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: Lett Appl Microbiol Asunto de la revista: MICROBIOLOGIA Año: 2006 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido