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Abstract

A counter-pressure-assisted capillary isotachophoresis method in combination with a sieving matrix and ionic spacer was used to perform in-line fluorescence in situ hybridization (FISH) of bacterial cells. A high concentration of sieving matrix (1.8% w/v HEC) was introduced at one end of the capillary, and the bacterial cells were suspended in the spacer electrolyte for injection. Using a 2 min injection with 18 psi counter-pressure, 50% of the cells injected into the capillary were hybridized with the fluorescently labeled oligonucleotide, and the excess unhybridized probe was separated from the hybridized cell–probe complexes in a two-stage ITP method. With an LOD (6.0 × 104 cells/mL) comparable with the CE analysis of a sample processed using an off-line FISH protocol, the total analysis time was reduced from 2.5 h to 30 min. Provided the appropriate probe is selected, this approach can be used for specific detection of bacterial cells in aqueous samples.

Item Type:	Article
Authors/Creators:	Phung, SC and Cabot, JM and Macka, M and Powell, SM and Guijt, RM and Breadmore, M
Keywords:	fluorescence in situ hybridization, bacterial cells,
Journal or Publication Title:	Analytical Chemistry
Publisher:	Amer Chemical Soc
ISSN:	0003-2700
DOI / ID Number:	https://doi.org/10.1021/acs.analchem.7b00598
Copyright Information:	Copyright 2017 American Chemical Society
Related URLs:	UTAS Profile: Phung, SC UTAS Profile: Cabot, JM Google Scholar: Macka, M UTAS Profile: Macka, M UTAS Profile: Powell, SM Google Scholar: Guijt, RM UTAS Profile: Guijt, RM Google Scholar: Breadmore, M UTAS Profile: Breadmore, M
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