A new electro-optical approach to rapid assay of cell viability.
A new electro-optical (EO) approach was developed and applied to rapidly assay cell viability by using phage M13K07. Since phage M13K07 can replicate only in living bacteria and cannot replicate in the presence of inhibitors, the difference between the EO signals obtained in the presence and absence of the phage can be used as an important factor for evaluating cell viability. Variation in the electrophysical parameters of Escherichia coli XL-1 during its interaction with phage M13K07 was studied under exposure of the cells to various inhibitors of cellular metabolism. Significant changes in the EO signal were found during incubation of living E. coli cells with phage M13K07. At the same time, no changes were recorded during cell incubation with the phage after pretreatment of E. coli XL-1 cells with sodium azide, carbonyl cyanide 3-chlorophenyl hydrazone, chloramphenicol, and kanamycin. This finding can be explained by the decrease in the number of living cells in the culture after preliminary incubation with the chemical agents, and it was confirmed by colony counts by conventional plating onto solid LB medium before and after treatment of the cells with the inhibitors. The EO approach can be used as a rapid method for evaluation of the inhibitory effects of various chemical agents and drugs, and it has the potential for the study of the molecular mechanisms underlying cell death.
Guliy
Olga I
OI
Institute of Biochemistry and Physiology of Plants and Microorganisms, RAS, 13 Prospekt Entuziastov, Saratov 410049, Russian Federation.
Bunin
Victor D
VD
O'Neil
Daniel
D
Ivnitski
Dmitri
D
Ignatov
Oleg V
OV
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D017104:Bacteriophage M13 / Q000254:growth & development; D002701:Chloramphenicol / Q000494:pharmacology; D015169:Colony Count, Microbial; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000502:physiology* / Q000821:virology; D007612:Kanamycin / Q000494:pharmacology; D050296:Microbial Viability / Q000187:drug effects*; D055095:Optics and Photonics
D000900:Anti-Bacterial Agents; D007612:Kanamycin; D002701:Chloramphenicol