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Isotachophoretic fluorescence in situ hybridization of intact bacterial cells
journal contribution
posted on 2023-05-19, 09:42 authored by Phung, SC, Cabot, JM, Miroslav MackaMiroslav Macka, Shane PowellShane Powell, Rosanne Guijt, Michael BreadmoreMichael BreadmoreA 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.
History
Publication title
Analytical ChemistryVolume
89Issue
12Pagination
6513-6520ISSN
0003-2700Department/School
School of Natural SciencesPublisher
Amer Chemical SocPlace of publication
1155 16Th St, Nw, Washington, USA, Dc, 20036Rights statement
Copyright 2017 American Chemical SocietyRepository Status
- Restricted