PMID- 28528550
OWN - NLM
STAT- MEDLINE
DCOM- 20190306
LR  - 20190306
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 89
IP  - 12
DP  - 2017 Jun 20
TI  - Isotachophoretic Fluorescence in Situ Hybridization of Intact Bacterial Cells.
PG  - 6513-6520
LID - 10.1021/acs.analchem.7b00598 [doi]
AB  - 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 10(4) 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.
FAU - Phung, Sui C
AU  - Phung SC
FAU - Cabot, Joan M
AU  - Cabot JM
FAU - Macka, Mirek
AU  - Macka M
AUID- ORCID: 0000-0002-6792-2574
FAU - Powell, Shane M
AU  - Powell SM
FAU - Guijt, Rosanne M
AU  - Guijt RM
AD  - Microfluidic Chipshop, GmbH, Stockholmer Strasse, Jena 07747, Germany.
FAU - Breadmore, Michael
AU  - Breadmore M
AUID- ORCID: 0000-0001-5591-4326
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170606
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Electrolytes)
SB  - IM
MH  - Cells, Cultured
MH  - Electrolytes/chemistry
MH  - Escherichia coli/*cytology
MH  - *In Situ Hybridization, Fluorescence
MH  - *Isotachophoresis
MH  - Pseudomonas aeruginosa/*cytology
EDAT- 2017/05/23 06:00
MHDA- 2019/03/07 06:00
CRDT- 2017/05/23 06:00
PHST- 2017/05/23 06:00 [pubmed]
PHST- 2019/03/07 06:00 [medline]
PHST- 2017/05/23 06:00 [entrez]
AID - 10.1021/acs.analchem.7b00598 [doi]
PST - ppublish
SO  - Anal Chem. 2017 Jun 20;89(12):6513-6520. doi: 10.1021/acs.analchem.7b00598. Epub 
      2017 Jun 6.