A protein-based oxygen biosensor for high-throughput monitoring of cell growth and cell viability.
Fluorescently labeled hemocyanin has been previously proposed as an oxygen sensor. In this study, we explored the efficacy of this biosensor for monitoring the biological oxygen consumption of bacteria and its use in testing bacterial cell growth and viability of Escherichia coli, Pseudomonas aeruginosa, Paracoccus denitrificans, and Staphylococcus simulans. Using a microwell plate, the time courses for the complete deoxygenation of samples with different initial concentrations of cells were obtained and the doubling times were extracted. The applicability of our fluorescence-based cell growth assay as an antibacterial drug screening method was also explored. The results provide a proof-of-principle for a simple, quantitative, and sensitive method for high-throughput monitoring of prokaryotic cell growth and antibiotic susceptibility screening.
Strianese
Maria
M
Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.
Zauner
Gerhild
G
Tepper
Armand W J W
AW
Bubacco
Luigi
L
Breukink
Eefjan
E
Aartsma
Thijs J
TJ
Canters
Gerard W
GW
Tabares
Leandro C
LC
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001419:Bacteria / Q000166:cytology* / Q000254:growth & development; D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development; D006433:Hemocyanins; D008826:Microbial Sensitivity Tests; D010100:Oxygen / Q000032:analysis*; D010231:Paracoccus denitrificans / Q000166:cytology / Q000254:growth & development; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000254:growth & development; D013210:Staphylococcus / Q000166:cytology / Q000254:growth & development
D000900:Anti-Bacterial Agents; D006433:Hemocyanins; D010100:Oxygen