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Isotachophoretic fluorescence in situ hybridization of intact bacterial cells

Citation

Phung, SC and Cabot, JM and Macka, M and Powell, SM and Guijt, RM and Breadmore, M, Isotachophoretic fluorescence in situ hybridization of intact bacterial cells, Analytical Chemistry, 89, (12) pp. 6513-6520. ISSN 0003-2700 (2017) [Refereed Article]

Copyright Statement

Copyright 2017 American Chemical Society

DOI: doi:10.1021/acs.analchem.7b00598

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 Details

Item Type:Refereed Article
Keywords:fluorescence in situ hybridization, FISH
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Instrumental Methods (excl. Immunological and Bioassay Methods)
Objective Division:Health
Objective Group:Public Health (excl. Specific Population Health)
Objective Field:Behaviour and Health
Author:Phung, SC (Ms Sui Phung)
Author:Cabot, JM (Mr Joan Cabot Canyelles)
Author:Macka, M (Professor Mirek Macka)
Author:Powell, SM (Dr Shane Powell)
Author:Guijt, RM (Dr Rosanne Guijt)
Author:Breadmore, M (Professor Michael Breadmore)
ID Code:120105
Year Published:2017
Deposited By:Chemistry
Deposited On:2017-08-10
Last Modified:2017-09-07
Downloads:0

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