alexa Staining of Escherichia coli for flow cytometry: influx and efflux of ethidium bromide.
Genetics & Molecular Biology

Genetics & Molecular Biology

Journal of Molecular Biomarkers & Diagnosis

Author(s): Jernaes MW, Steen HB

Abstract Share this page

Abstract In an attempt to develop procedures for nucleic acid staining of bacteria for clinical routine assays, the uptake of ethidium bromide (EB) in wild-type Escherichia coli was studied using flow cytometry. Phosphate-buffered saline (PBS) containing EDTA or Tris significantly increased the net uptake of EB compared to PBS only. However, in the majority of the cells, the net uptake reached a constant level that was only a few percent of that of fully permeabilized cells, apparently due to the activity of a metabolically driven efflux pump. When cells were exposed to cold shock (0 degrees C for 30 min) in the presence of Tris or EDTA, the net uptake of dye was similar to that of fully permeabilized cells, whereas it was about half that value in PBS. When cold shock was given in growth medium, the cells split up into four subpopulations, with a net dye uptake ranging from that of fully permeabilized cells to less than 1\% of that value. As expected, metabolic inhibitors (Na-azide, 2-deoxy-D-glucose, and CCCP) reduced efflux activity. However, fluorescence of metabolically inhibited cells never exceeded more than about half the value of that of dead cells, possibly reflecting conformational changes in DNA structure as a result of cell death. This article was published in Cytometry and referenced in Journal of Molecular Biomarkers & Diagnosis

Relevant Expert PPTs

Relevant Speaker PPTs

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

[email protected]

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

[email protected]

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001 Extn: 9042

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords