Total internal reflection fluorescence microscopy of intraflagellar transport in Tetrahymena thermophila.
Total internal reflection fluorescence microscopy of intraflagellar transport in Tetrahymena thermophila.
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DOI：
10.1016/bs.mcb.2015.01.001
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发表时间：
2015
影响因子：
--
通讯作者：
Gaertig J
中科院分区：
文献类型：
--
作者：
Jiang YY;Lechtreck K;Gaertig J
活体成像已成为研究纤毛蛋白的有力工具。嗜热四膜虫是一种成熟的纤毛虫模型，具有成熟的遗传和生化手段，但其体积大、形状复杂、纤毛数量多且短而重叠，给纤毛蛋白的实时成像带来了挑战。在这里，我们描述了一种结合镍离子麻痹纤毛和全内反射显微镜的方法，用于实时成像四膜虫纤毛内的荧光蛋白。利用这种方法，我们定量地记录了四膜虫鞭毛内的运输。
Live imaging has become a powerful tool in studies of ciliary proteins. Tetrahymena thermophila is an established ciliated model with well developed genetic and biochemical approaches, but its large size, complex shape and the large number of short and overlapping cilia, have made live imaging of ciliary proteins challenging. Here we describe a method that combines paralysis of cilia by nickel ions and total internal reflection microscopy for live imaging of fluorescent proteins inside cilia of Tetrahymena. Using this method, we quantitatively documented the intraflagellar transport in Tetrahymena.