Flow cytometric characterization and sorting of cultured Brassica napus microspores. 1991

C Deslauriers, and A D Powell, and K Fuchs, and K P Pauls
Department of Crop Science, University of Guelph, Ontario, Canada.

A flow cytometric procedure was used to characterize and sort embryogenic Brassica napus microspore cultures. Embryogenic microspores continued to increase in size throughout the culture period and fluoresced when stained with fluoresecein diacetate (FDA). However, most of the cells in culture (greater than 95%) lost their viability over the culture period and the correlation between the percentage viable cells in a culture and the productivity of the culture was not significant until day 3. By sorting large fluorescent cells on day 1 and day 3 of the culture period populations of cells that were 7-18-fold more embryogenic than sorted mixed cells were obtained. The flow cytometric procedure would be useful for rapidly assessing the effects of a large variety of culture and media conditions on embryogenesis is rapeseed microspore cultures. Furthermore, the populations of potentially embryogenic microspores isolated by flow cytometry might be useful for studies of the early cellular and molecular changes that occur during androgenesis in rapeseed.

UI MeSH Term Description Entries
D001937 Brassica A plant genus of the family Cruciferae. It contains many species and cultivars used as food including cabbage, cauliflower, broccoli, Brussel sprouts, kale, collard greens, MUSTARD PLANT; (B. alba, B. junica, and B. nigra), turnips (BRASSICA NAPUS) and rapeseed (BRASSICA RAPA). Broccoli,Brussel Sprout,Cabbage,Cauliflower,Collard Green,Kale,Cabbages,Collard Greens
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005452 Fluoresceins A family of spiro(isobenzofuran-1(3H),9'-(9H)xanthen)-3-one derivatives. These are used as dyes, as indicators for various metals, and as fluorescent labels in immunoassays. Tetraiodofluorescein

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