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Going for Organelles
The suitability of protoplasts for analysis of cellular organelles such as the nucleus, is compromised by their acentric position in the cell. Because of this, even the regular movement of protoplasts in the flow will not guarantee the same position of all the organelles to be measured with respect to the excitation light beam and optical detector system. Moreover, light may scatter on internal components of protoplasts and disturb the analysis. It is then logical to isolate the organelles and...
Plant Pathogens
Although not explored extensively, FCM can be used as a convenient and rapid tool to detect plant pathogens e.g. viruses, bacteria, and fungi , and to assess their viability see Chapter 9 . As in other applications, FCM excels in its ability to analyze large population samples in a short time and to discriminate subpopulations. Identification of specific viruses, bacteria and fungal spores can be achieved after immunofluorescent staining of a pathogen in a crude plant extract Chitarra and van...
Ploidy Levels
Chromosome counting in dividing cells is an unambiguous way to determine the ploidy, or the number of basic chromosome sets in cell nuclei. However, this is time consuming and tissues containing dividing cells may not be readily available. Hence, alternatives which do not require mitotically-active tissues and chromosome preparations were always sought. But, none of the early options, such as the estimation of leaf stomata density and size and pollen grain size, were found to be sufficiently...
Secondary Metabolites
Plants can produce and accumulate usually in vacuoles a wide range of secondary metabolites with protective or signaling functions. They may be a focal point of biotechnology processes and their presence, distribution, and concentration may be analyzed with FCM Yanpaisan et al. 1999 . The most comprehensive data apparently concern Catharanthus roseus Apocynaceae where FCM has been used to quantify both alkaloid Brown et al. 1983 and anthocyanin Hall and Yeoman 1987 content in isolated...
The Growth of Multiparameter Flow Cytometry
In the early 1970s, the group at Los Alamos led the way in implementation of practical multiparameter flow cytometers their larger instruments, with droplet sorting capability, combined two-color fluorescence measurements with measurements of Coulter volume and thanks to the contributions of Paul Mullaney, Gary Salzman, and others light scattering at several angles Mullaney et al. 1969 Salz-man et al. 1975a, 1975b Steinkamp et al. 1973 . The cytometers were interfaced to Digital Equipment...