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Isotachophoretic Fluorescence in Situ Hybridization of Intact Bacterial Cells.
- Source :
-
Analytical Chemistry . 6/20/2017, Vol. 89 Issue 12, p6513-6520. 8p. - Publication Year :
- 2017
-
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 x 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. [ABSTRACT FROM AUTHOR]
- Subjects :
- *ISOTACHOPHORESIS
*ELECTROPHORESIS
*BACTERIAL cells
*ELECTROLYTES
*OLIGONUCLEOTIDES
Subjects
Details
- Language :
- English
- ISSN :
- 00032700
- Volume :
- 89
- Issue :
- 12
- Database :
- Academic Search Index
- Journal :
- Analytical Chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 123832943
- Full Text :
- https://doi.org/10.1021/acs.analchem.7b00598