Quantification of bacterial cells based on autofluorescence on a microfluidic platform

被引:37
作者
Bao, Ning [1 ]
Jagadeesan, Balarnurugan [2 ]
Bhunia, Arun K. [2 ]
Yao, Yuan [2 ]
Lu, Chang [1 ,3 ,4 ]
机构
[1] Purdue Univ, Dept Agr & Biol Engn, W Lafayette, IN 47907 USA
[2] Purdue Univ, Dept Food Sci, Mol Food Microbiol Lab, W Lafayette, IN 47907 USA
[3] Purdue Univ, Birck Nanotechnol Ctr, W Lafayette, IN 47907 USA
[4] Purdue Univ, Bindley Biosci Ctr, W Lafayette, IN 47907 USA
关键词
microfluidics; electrophoresis; laser-induced fluorescence; autofluorescence; electrical lysis; enumeration;
D O I
10.1016/j.chroma.2007.12.048
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Bacterial counts provide important information during the processes such as pathogen detection and hygiene inspection and these processes are critical for public health and food/pharmaceutical production. In this study, we demonstrate the quantification of the number of bacterial cells based on the autofluorescence from the cell lysate on a microfluidic chip. We tested three model pathogenic bacteria (Listeria monocytogenes F4244, Salmonella Enteritidis PT1 and Escherichia coli O157:H7 EDL 933). In the experiment, a plug of similar to 150 pL containing lysate from 240 to 4100 cells was injected into a microfluidic channel with downstream laser-induced fluorescence detection under electrophoresis conditions. We found that the autofluorescence intensity increased with the number of cells almost linearly for all three bacteria. The autofluorescence remained a single peak when the cell lysate contained a mixture of different bacterial species. We also demonstrate a simple microfluidic device that integrates entrapment and electrical lysis of bacterial cells with fluorescence detection. Such a device can carry out the quantification of bacterial cells based on lysate autofluorescence without off-chip procedures. This study offers a simple and fast solution to on-chip quantification of bacterial cells without labeling. We believe that the method can be extended to other bacterial species. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:153 / 158
页数:6
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