Title
Flow cytometric enumeration of bacteria using TO-PRO®-3 iodide as a single-stain viability dye Flow cytometric enumeration of bacteria using TO-PRO®-3 iodide as a single-stain viability dye
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences. Pharmacy
Publication type
article
Publication
New York :Elsevier ,
Subject
Chemistry
Biology
Pharmacology. Therapy
Source (journal)
Journal of laboratory automation / Society for Laboratory Automation and Screening. - New York, 2011, currens
Volume/pages
19(2014) :6 , p. 555-561
ISSN
2211-0682
ISI
000345468000005
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Quantification of bacteria using conventional viable plate counting (VPC) is labor-intensive and time-consuming. Flow cytometry (FCM) can be proposed as a faster alternative. This study aimed to develop a flow cytometric, single-stain approach using TO-PRO®-3 iodide (TP3) for the quantification of Staphylococcus aureus, Escherichia coli, and Bacillus subtilis cells. Live or dead bacterial suspensions were stained with TP3 and analyzed using a FACSCalibur flow cytometer. After optimization of staining parameters and instrument settings, an excellent separation of viable and dead cells was achieved for all species. The quantitative performance of the technique was assessed by analyzing serial dilutions of bacterial suspensions using FCM and VPC. A highly linear correlation (r2 > 0.99) was observed between the colony forming units (CFU)/mL as determined by FCM and by VPC over a concentration range of about 104 to 108 CFU/mL. As such, FCM quantification of viable bacteria using TP3 can be considered as an accurate and reliable alternative for VPC. The monostain procedure is easy to apply and cost-effective, and it allows bacterial enumeration in a broad variety of samples.
Full text (open access)
https://repository.uantwerpen.be/docman/irua/e0e73f/9053.pdf
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