Magnetic Separation of Phagosomes of Defined Age from Tetrahymena thermophila |
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Authors: | CHRISTIAN VOSKÜHLER ARNO TIEDTKE |
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Institution: | Institute for General Zoology and Genetics, University of Münster, Schlossplatz 5, 48149 Münster, Germany |
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Abstract: | ABSTRACT. We describe a new mass isolation procedure for both pure and stage-specific phagosomes from Tetrahymena thermophila . We prepared magnetic iron dextran particles about 1 μm in diameter to label the phagosomes. The oral apparatus of the cells concentrated these particles so readily that after 1 min the majority of the cells had formed a single phagosome. A short wash removed non-ingested particles, enabling us to follow the age-dependent changes of a single labeled phagosome through the cell. Phagosomes of different ages, including very young and nascent phagosomes, were removed easily from the non-magnetic cell debris of mechanically homogenized cells by means of a permanent magnet. The isolated phagosomes are pure as tested by enzymatic assays and light and electron microscopy. Since the yield of pure phagosomes of all ages is high (∼ 90%), this method could be generally applied for phagosome isolation from ciliates. |
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Keywords: | Endosomes isolation-method |
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