Ref ID : 7288
Christian Vosskuhler and Arno Tiedtke; Magnetic Separation of Phagosomes of Defined Age from Tetrahymena thermophila. J.Eukaryot.Microbiol. 40(5):556-562, 1993
Reprint
In File
Notes
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.