Individual Escherichia coli cells studied from light scattering with the scanning flow cytometer

• Published in: Cytometry (New York, N.Y.) N.Y.), 2000-09, Vol.41 (1), p.41-45
• Main Authors: Shvalov, Alexander N., Soini, Juhani T., Surovtsev, Ivan V., Kochneva, Galina V., Sivolobova, Galina F., Petrov, Alexander K., Maltsev, Valeri P.
• Format: Article
• Language: English
• Subjects: differential cross section; Escherichia coli - cytology; Escherichia coli - isolation & purification; Flow Cytometry - instrumentation; Flow Cytometry - methods; growth phases; Microbiological Techniques; microorganism identification; Scattering, Radiation; T‐matrix method
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/1097-0320(20000901)41:1<41::AID-CYTO6>3.0.CO;2-N

Background Flow cytometry is a powerful tool for the analysis of individual particles in a flow. Differential light scattering (an indicatrix) was used for many years to obtain morphologic information about microorganisms. The indicatrices play the same role for individual particle recognition as a spectrum for substance characterization. We combined two techniques to analyze the indicatrix of the cells for the purpose of developing a database of light‐scattering functions of cells. Methods The scanning flow cytometer (SFC) allows the measurement of the entire indicatrix of individual particles at polar angles ranging from 5° to 100°. In this work, light‐scattering properties of Escherichia coli have been studied both experimentally and theoretically with the SFC and the T‐matrix method, respectively. The T‐matrix method was used because of the nonspherical shape of E. coli cells, which were modeled by a prolate spheroid. Results The indicatrices of E. coli cells were stimulated with T‐matrix method at polar angles ranging from 10° to 60°. The absolute cross‐section of light scattering of E. coli has been determined comparing the cross section of polystyrene particles modeled by a homogeneous sphere. The E. coli indicatrices were compared for logarithmic and stationary phases of cell growth. Conclusions The indicatrices of E. coli were reproducible and could be used for identification of these cells in biologic suspensions. The angular location of the indicatrix minimum can be used in separation of cells in logarithmic and stationary phases. To use effectively the indicatrices for that purpose, the light‐scattering properties of other microorganisms have to be studied. Cytometry 41:41–45, 2000 © 2000 Wiley‐Liss, Inc.