Recent advances of flow cytometry in fundamental and applied microbiology

Abstract

This review focuses on the recent applications of flow cytometry (FCM) in microbiological research (1987-mid 1992). It tries to give a scope of the important breakthroughs which occurred in this field during this period. The technical difficulties of microorganism analysis by flow cytometry is briefly appraised. The significance and the limits of the different microbial cell parameters attainable by flow analyses are systematically evaluated: light scatter for cell size and structure, fluorescence measurements for quantification of cellular components, microbial antigen detection and cell physiological activity estimation. Emphasis is given on the new technological advances which appeared in the last two years. The second part of the review is devoted to the analysis of the usefulness of flow cytometric approach in the different fields of microbiology: fundamental studies in microbial physiology, differentiation, microbial ecology and aquatic sciences, medical microbiology, parasitology, microbial pharmacology and biotechnology.

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

“…Although some nonspecific identification can always occur, detection of 1 positive cell in a background of 10000 negative cells is possible. Fouchet et al (1993) reviews this area of research that, to this moment, has not involved any studies with natural planktonic bacteria, although immunofluorescence combined with flow cytometry was recently used to assess selective removal of natural bacterial strains by protistan grazers (Frette and del Giorgio, unpublished). Fluorescent in situ hybridization (FISH) has also the potential for being very useful in the near future.…”

“…The current explosion in the use of FC in ecological studies has been in part fueled by the availability of new nuclear acid stains together with powerful, sensitive and relatively cheap benchtop flow cytometers. Few areas of research or few techniques have had such an amount of review papers and books in relatively few years: Darzynkiewicz and Crissman (1990), Ormerod (1994), Lloyd (1993), Fouchet et al (1993), Troussellier et al (1993), Methods in Cell Biology (1994), Shapiro (1995), Davey and Kell (1996), Porter et al (1997), Davey et al (1999), Collier and Campbell (1999), etc. However, and with the exception of the Trousellier et al (1993) paper, and small sections in the complete reviews of Davey and Kell (1996) and Collier and Campbell (1999), little has been published on the application of flow cytometry to natural planktonic bacteria, an area that has flourished after the papers of Li et al (1995), del Giorgio et al (1996) and Marie et al (1997) that independently realized the potential of the blue-light excitable stains marketed by Molecular Probes.…”

Flow Cytometric Assessment of Viability of Lactic Acid Bacteria

“…It is used in counting the total number of bacteria and in detecting specific strains by 16S rRNA se-quence or by antigen expression. It is also used for characterizing and quantifying cellular physiological parameters such as DNA content, enzyme activity, respiration, membrane potential, intracellular pH, and membrane integrity (11,16,27,34,35).…”

Accurate quantitation ofLeishmania infection in cultured cells by flow cytometry

“…While the FCM technique has found a broad range of applications in bacteriology, its use in parasitology has been largely restricted to the study of extracellular parasites (19). One exception is the group of the hemoparasites which, residing in nonnucleated red blood cells, can be specifically labeled with fluorescent DNA-binding dyes (20 -23).…”