METHOD AND APPARATUS FOR LABELING PLANT CELL NUCLEUS

摘要

PURPOSE:To introduce 2 kinds of fluorescent dyes consisting of an acridine group and phenyl indole group into various plant protoplast cells in a short period of time by impressing electric pulses to the cells. CONSTITUTION:The grown leave of, for example, celery (Apium graveolens Linn. var dulce DC) is sterilized and the thin skin thereof is stripped. Cell wall decomposition enzyme is put into 0.6M mannitol liquid adjusted to 5.7pH to form the bare cells in a cell 1. The cells are sieved by a nylon net 4 and are transferred to a dyeing cell 2. The cells are thereafter cleaned by a sterilizing liquid. The suspension thereof is adjusted to the concn. of Acridine orange (AO) 50-250muM and about 10 cells/ml cell concn. The cells are labeled in 3-5min when 300-500V pulse voltage is impressed at 1-3 times from an electric pulse generator 9 via electrodes to the cells. The nuclei and cytoplasma are then observed by a fluorescent microscope. The fluorescent wavelength region of the AO glows yellow at 520nm and is, therefore, easily identifiable. The labeled bare cells of another leave are prepd. by another apparatus of the same type. The unnecessary enzyme and dyes are thereafter washed away and the cells are sent to a fusion cell 3 where the cell fusion is executed.