Jiang Yu-Yang, Lechtreck Karl, Gaertig Jacek
Department of Cellular Biology, University of Georgia, Athens, GA, USA.
Methods Cell Biol. 2015;127:445-56. doi: 10.1016/bs.mcb.2015.01.001. Epub 2015 Feb 14.
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.
实时成像已成为研究纤毛蛋白的强大工具。嗜热四膜虫是一种成熟的纤毛模型，具有完善的遗传学和生物化学方法，但其体积大、形状复杂以及大量短而重叠的纤毛，使得对纤毛蛋白进行实时成像具有挑战性。在此，我们描述了一种结合镍离子使纤毛麻痹和全内反射显微镜技术的方法，用于嗜热四膜虫纤毛内荧光蛋白的实时成像。使用这种方法，我们定量记录了嗜热四膜虫中的鞭毛内运输。
