A new electro-optical approach to rapid assay of cell viability

被引:6
作者
Guliy, Olga I.
Bunin, Victor D.
O'Neil, Daniel
Ivnitski, Dmitri
Ignatov, Oleg V.
机构
[1] RAS, Inst Biochem & Physiol Plants & Microorganisms, Saratov 410049, Russia
[2] State Res Ctr Appl Microbiol & Biotechnol, Obolensk, Russia
[3] CRADA Int Inc, Norcross, GA USA
[4] New Mexico Inst Min & Technol, Inst Engn Res & Appl, Albuquerque, NM 87106 USA
[5] Univ New Mexico, Dept Chem & Nucl Engn, Albuquerque, NM 87131 USA
关键词
Escherichia coli XL-1; orientational spectrum; phage M13K07; cell viability;
D O I
10.1016/j.bios.2007.06.008
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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. (c) 2007 Published by Elsevier B.V.
引用
收藏
页码:583 / 587
页数:5
相关论文
共 25 条
[1]  
Adams M. H., 1959, BACTERIOPHAGES
[2]   DIELECTRIC ENERGY OF ORIENTATION IN DEAD AND LIVING CELLS OF SCHIZOSACCHAROMYCES-POMBE - FITTING OF EXPERIMENTAL RESULTS TO A THEORETICAL-MODEL [J].
ASENCOR, FJ ;
SANTAMARIA, C ;
IGLESIAS, FJ ;
DOMINGUEZ, A .
BIOPHYSICAL JOURNAL, 1993, 64 (05) :1626-1631
[3]   Assessment of viability of microorganisms employing fluorescence techniques [J].
Breeuwer, P ;
Abee, T .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 2000, 55 (1-3) :193-200
[4]   Determination of cell structures, electrophysical parameters, and cell population heterogeneity [J].
Bunin, VD ;
Voloshin, AG .
JOURNAL OF COLLOID AND INTERFACE SCIENCE, 1996, 180 (01) :122-126
[5]   Electrooptical analysis of the Escherichia coli-phage interaction [J].
Bunina, VD ;
Ignatov, OV ;
Guliy, OI ;
Zaitseva, IS ;
O'Nell, D ;
Ivnitski, D .
ANALYTICAL BIOCHEMISTRY, 2004, 328 (02) :181-186
[6]   Filamentous phage infection: Required interactions with the TolA protein [J].
Click, EM ;
Webster, RE .
JOURNAL OF BACTERIOLOGY, 1997, 179 (20) :6464-6471
[7]   Interaction of the globular domains of pIII protein of filamentous bacteriophage fd with the F-pilus of Escherichia coli [J].
Deng, LW ;
Malik, P ;
Perham, RN .
VIROLOGY, 1999, 253 (02) :271-277
[8]  
Gerhardt P., 1981, Manual of methods for general bacteriology
[9]   A unified resistor-capacitor model for impedance, dielectrophoresis, electrorotation, and induced transmembrane potential [J].
Gimsa, J ;
Wachner, D .
BIOPHYSICAL JOURNAL, 1998, 75 (02) :1107-1116
[10]  
Gottschalk G., 1979, BACTERIAL METABOLISM