A flow cytometric approach for characterization and differentiation of bacteria during microbial processes

authored by: S. Müller, A. Lösche, T. Bley, T. Scheper
Abstract: The analysis of growing or resting bacterial populations by flow cytometry offers several advantages over traditional methods for determining mean-value parameters. This method has been applied here to measure both the distribution of single-cell fluorescence intensity and the light-scatter behaviour of the methylotrophical strains of Methylobacterium rhodesianum MB126 and Methylocystis GB25 as well as Pseudomonas fluorescens and a strain isolated from the soil. The four different bacterial populations were analysed concerning the DNA and the poly-3-hydroxybutyrate (PHB) content. A new cell-preservation method is presented. Optimized staining methods for each strain were developed in detail, in two cases DNA had to be dehybridized before staining with a mixture of mithramycin/ethidium bromide. Nile red is used for detecting PHB. Both stains were excited by an argonion laser at 488 nm; fluorescence emission for mithramycin/ethidium bromide was measured from 520 nm and for Nile red from 600 nm onwards. It is shown that changes in the DNA content and in the forward-light-scattering behaviour of the bacterial strains chosen were measurable. These changes could be related to different cultivation conditions and correlated, in the case of strains that accumulate PHB, with alterations of that biopolymer content. In addition it was found that these methods provide a contribution to the differentiation of mixed bacterial populations.
External Organisation(s): University of Münster
Leipzig University
Type: Article
Journal: Applied Microbiology and Biotechnology
Volume: 43
Pages: 93-101
No. of pages: 9
ISSN: 0175-7598
Publication date: 04.1995
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Biotechnology, Applied Microbiology and Biotechnology
Electronic version(s): https://doi.org/10.1007/BF00170629 (Access: Unknown)

BibTeX

@article{ab0eb51ebf384ea4b8adca5cb03e9809,
title = "A flow cytometric approach for characterization and differentiation of bacteria during microbial processes",
abstract = "The analysis of growing or resting bacterial populations by flow cytometry offers several advantages over traditional methods for determining mean-value parameters. This method has been applied here to measure both the distribution of single-cell fluorescence intensity and the light-scatter behaviour of the methylotrophical strains of Methylobacterium rhodesianum MB126 and Methylocystis GB25 as well as Pseudomonas fluorescens and a strain isolated from the soil. The four different bacterial populations were analysed concerning the DNA and the poly-3-hydroxybutyrate (PHB) content. A new cell-preservation method is presented. Optimized staining methods for each strain were developed in detail, in two cases DNA had to be dehybridized before staining with a mixture of mithramycin/ethidium bromide. Nile red is used for detecting PHB. Both stains were excited by an argonion laser at 488 nm; fluorescence emission for mithramycin/ethidium bromide was measured from 520 nm and for Nile red from 600 nm onwards. It is shown that changes in the DNA content and in the forward-light-scattering behaviour of the bacterial strains chosen were measurable. These changes could be related to different cultivation conditions and correlated, in the case of strains that accumulate PHB, with alterations of that biopolymer content. In addition it was found that these methods provide a contribution to the differentiation of mixed bacterial populations.",
author = "S. M{\"u}ller and A. L{\"o}sche and T. Bley and T. Scheper",
year = "1995",
month = apr,
doi = "10.1007/BF00170629",
language = "English",
volume = "43",
pages = "93--101",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "1",
}