Flow cytometric analysis to detect pathogens in bacterial cell mixtures using semiconductor quantum dots

被引:73
作者
Hahn, Megan A. [1 ]
Keng, Peter C. [2 ]
Krauss, Todd D. [1 ]
机构
[1] Univ Rochester, Dept Chem, Rochester, NY 14627 USA
[2] Univ Rochester, Med Ctr, Dept Radiat Oncol, Rochester, NY 14642 USA
关键词
D O I
10.1021/ac7018365
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Compared to a common green organic dye, semiconductor quantum dots (QDs) composed of CdSe/ZnS core/ shell bioconjugates display brighter fluorescence intensities, lower detection thresholds, and better accuracy in analyzing bacterial cell mixtures composed of pathogenic E. coli O157:H7 and harmless E. coli DH5 alpha using flow cytometry. For the same given bacterial mixture, QDs display fluorescence intensity levels that are similar to 1 order of magnitude brighter compared to the analogous experiments that utilize the standard dye fluorescein isothiocyanate. Detection limits are lowest when QDs are used as the fluorophore label for the pathogenic E. coli O157:H7 serotype: limits of 1% O157:H7 in 99% DH5a. result, corresponding to 10(6) cells/mL, which is comparable to other developing fluorescence-based techniques for pathogen detection. Finally, utilizing QDs to label E. coli O157: H7 in cell mixtures results in greater accuracy and more closely approaches the ideal fluorophore for pathogen detection using flow cytometry. With their broader absorption spectra and narrower emission spectra than organic dyes, QDs can make vast improvements in the field of flow cytometry, where single-source excitation and simultaneous detection of multicolor species without complicating experimental setups or data analysis is quite advantageous for analyzing heterogeneous cell mixtures, both for prokaryotic pathogen detection and for studies on eukaryotic cell characteristics.
引用
收藏
页码:864 / 872
页数:9
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