A reinvestigation of cross-sections of cilia.
A reinvestigation of cross-sections of cilia.
复制标题
对纤毛的横截面的重新研究。
DOI：
10.1083/jcb.37.3.825
复制
发表时间：
1968-06
影响因子：
7.8
通讯作者：
Allen, R D
中科院分区：
文献类型：
--
作者：
Allen, R D
材料与方法以梨形囊膜虫（Stetrahymena pyriformis）W株为材料，进行了实验研究。培养程序和电子显微镜制备的细节在别处描述（艾伦，1967）。可能，与其他人在类似研究中通常采用的方法相比，固定方法中最显著的差异是使用戊二醛，然后使用OsO 4（Sabatini等人，1963)-而不是单独的OsO 4-和相对短的固定和脱水时间。在Porter-Blum MT-2超薄切片机上用金刚石刀切割切片，然后无支撑地安装在铜网格上。使用醋酸双氧铀（沃森，1958）和柠檬酸铅（Reynolds，1963）进行染色。然后在Philips EM 200或RCA EMU 3F电子显微镜中检查切片。为了增强纤毛照片中任何径向对称结构的密度，使用Markham等人首次应用于电子显微照片的旋转技术。（1963年）。为此目的，选择了特别有利的纤毛图像。然而，正如Ledbetter和Porter（1964）和Ringo（1967 a）所指出的，由于圆形对于使用这种技术是必不可少的，并且由于许多显示精细细节的纤毛有些椭圆形，这是切片过程中压缩的结果，因此采用了对Markham旋转技术的修改。不是将放大的图像投影在水平表面上，而是构造了一个可以倾斜的画架，然后将放大器产生的图像投影到由倾斜的画架形成的倾斜平面上。当这个倾斜的画架朝向
MATERIALS AND METHODSTetrahymena pyriformis, strain W, was used in this investigation. The culturing procedure and details of preparation for electron microscopy are described elsewhere (Allen, 1967). Probably, the most significant difference in the fixation procedure as compared with the procedures usually employed by others in similar investigations is the use of glutaraldehyde followed by OsO4 (Sabatini et al., 1963)-instead of Os04 alone-and relatively short fixation and dehydration times. Sections were cut on a Porter-Blum MT-2 ultramicrotome with a diamond knife and then mounted unsupported on copper grids. Staining was achieved with uranyl acetate (Watson, 1958) followed by lead citrate (Reynolds, 1963). The sections were then examined in a Philips EM 200 or RCA EMU 3F electron microscope.For the purpose of enhancing the density of any radially symmetrical structures in the photographs of cilia, use was made of the rotation technique first applied to electron micrographs by Markham et al.(1963). Particularly favorable images of cilia were selected for this purpose. However, since circularity is essential for the use of this technique, as has been pointed out by Ledbetter and Porter (1964) and Ringo (1967 a), and since many of the cilia showing fine detail were somewhat elliptical, as a result of compression during sectioning, a modification of the Markham rotation technique was employed. Instead of projecting the enlarged image on a horizontal surface, an easel that could be tilted was constructed and the image produced by the enlarger was then projected onto the inclined plane formed by the tilted easel. When this tilted easel is oriented