Abstract
Publisher Summary This chapter covers standard methods of enumeration of total bacterial numbers with epifluorescence microscopy. The advantages of flow cytometery over microscopic enumeration are discussed. Conversion factors used to estimate carbon biomass based on cell abundance and cell sizes are discussed, and also the Circulating Tumor Cell (CTC) assay, which identifies bacteria with highly active electron transport systems (ETS) are described. Bacterial abundance, cell size, biomass, and activity are among the most fundamental properties that characterize natural aquatic systems, and the measurement of these parameters is central to aquatic microbial ecology. Epifluorescence techiques are now routinely performed in most microbial laboratories, and image analysis has increasingly been used to determine cell size and other cellular properties. Flow cytometry offers the possibility of assessing not only total abundance with great speed and precision, but also properties of individual cells, such as size and metabolic activity. Cytometry also offers the possibility of physically separating, via cell sorting, subpopulations based on various cellular properties. Flow cytometry and image analysis combined with selective staining methods, permit determination of physiological condition of in situ bacterioplankton.
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