Flow cytometry is a powerful technology that allows for the simultaneous analysis of multiple attributes of cells or particles in a liquid medium. The first cytometer used was built during World War II, when [1] developed an equipment where particles flowed through the system to diffuse light through a lens, producing electrical signals sensed by a photodetector. The instrument could detect objects in the order of ~ 0.5 µm in diameter, and is recognized as the first flow cytometer used for observation of biological cells [2]. This would be possible to identify aerosols, bacteria that would possibly biological warfare agents as well as check the efficiency of gas mask filters against particles. In 1950, the same principle was applied to the detection and enumeration of blood cells. As hematology and cellular immunology, two biological areas, that drove the development of flow cytometry [3]. Later, with improved equipment and methods, this technique was adapted to other areas of biology, including the plant kingdom [4]. Already in 1973 the German botanist Friedrich Otto Heller used the Impulszytophotometrie (pulse cytophotometry in German). This scientist did not imagine that it has launched a new field of scientific research, which would later be called flow cytometry in plants.
In reference to [5] that developed a rapid and convenient method for the isolation of plant nuclei by cutting the same tissue in a lysis buffer consisting of a buffer to destroy the
cellular and nuclear membranes of the cell allowing the release of DNA. Since then, this has been the main and most reliable method of isolating nuclear plant in flow cytometry. Any type of sample can be analyzed because its particles (cells, nuclei, chromosomes, cell organelles, or other cell subparticles) are suspended and vary between 0.2 µm and 50 µm in size. Solid tissues must be disaggregated and suspended before flow cytometry analysis. The suspended particles are then placed into a flow cytometry device.
The studies on flow cytometry have used as base the plant tissue culture, including the regeneration of plants subjected to chromosome doubling, for detection of somaclonal variation in material micropropagated in various subcultures, viability of pollen grains, cell cycles and the determination of ploidy. This chapter presents results obtained through flow cytometry on plant tissue culture.
No comments:
Post a Comment