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Flow cytometry and plant protoplast cell biology
Author(s) -
Bergounioux Catherine,
Brown Spencer C.,
Petit Patrice X.
Publication year - 1992
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1992.tb04750.x
Subject(s) - protoplast , flow cytometry , biology , cell , cytoplasm , dna , organelle , plant cell , microbiology and biotechnology , biophysics , biochemistry , gene
Flow cytometry, which requires individualized or single cell preparations, is particularly adapted to studies of protoplast biology. It has been developed at three levels: descriptions of populations, measurement of cell function, and positive or negative selection of rare protoplast types. It enables measurement of several parameters in single protoplasts at a high rate of analysis. Without protoplast purification, excellent resolution of subpopulations is still possible. The quantitative data obtained on each protoplast normally have relative units but absolute calibration of intensity scale is available. Specific probes allow one to correlate multiparameter data on each cell. DNA, RNA, proteins, metabolites, organelles, virus and plasmid content of protoplasts have been described. The essential conceptual advantage of flow cytometry is the processing of heterogeneous material, typical of biological samples, but inadequately handled through overall mean values. Biological properties of protoplast subpopulations are followed using fluorescence intensity as a measure of DNA versus RNA or protein synthesis, cytoplasmic and nuclear antigens, membrane potentials, membrane viscosity, cell wall regeneration and pH.