A New Isolation Medium for Plant Organelles

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Abstract
A new cell-extraction medium was developed, under conditions such that physical injury was minimized, for the simultaneous isolation of plant nuclei, mitochondria, and chloroplasts in morphological forms closely resembling those in living cells. Comparisons were made of organelles isolated in buffered NaCl, sucrose, and specially developed formulations with the synthetic high-molecular-weight polymers Ficoll (polymerized form of sucrose, MW 400,000) and Dextran 40 (MW 26,000). The finally developed medium contained: sucrose (osmoticum); Tris-acetate (buffer); Ficoll, Dextran 40, and bovine serum albumin (structure preservatives); glutathione, cysteine, ascorbic acid, or mercaptoethanol (reducing agents); and magnesium acetate and manganese acetate (activity preservatives). The properties of the SFDP medium are such that nuclei, chloroplasts, and mitochondria resemble their forms in living cells; mitochondria exhibit dynamic pleomorphic changes in form similar to those occurring in living cells and hair cells immersed in the medium can be cut open while cytoplasmic streaming continues during and after the incision.