Individual microtubules viewed by immunofluorescence and electron microscopy in the same PtK2 cell.
Individual microtubules viewed by immunofluorescence and electron microscopy in the same PtK2 cell.
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DOI：
10.1083/jcb.77.3.r27
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发表时间：
1978-06
期刊：
影响因子：
--
通讯作者：
Weber K
中科院分区：
文献类型：
--
作者：
Osborn M;Webster RE;Weber K
PtK 2细胞在金网格上生长，并在微管稳定缓冲液中用Triton X-100处理。用戊二醛固定所得细胞骨架，并使用单特异性微管蛋白抗体进行间接免疫荧光程序。首先通过荧光显微镜检查网格，并记录荧光细胞质微管的显示。然后用醋酸双氧铀对网格进行染色，并通过电子显微镜记录纤维结构的显示。因此，可以在相同的细胞骨架内比较光镜和电镜下细胞质微管结构的显示。结果表明，光学显微镜中的荧光纤维与电子显微镜中直径约550 A的不间断纤维直接对应。这是报道的由两层抗体分子围绕其圆周装饰的单个微管的直径。因此，在最佳条件下，免疫荧光显微镜可以显示单个微管。
PtK2 cells were grown on gold grids and treated with Triton X-100 in a microtubule stabilizing buffer. The resulting cytoskeletons were fixed with glutaraldehyde and subjected to the indirect immunofluorescence procedure using monospecific tubulin antibodies. Grids were examined first by fluorescence microscopy, and the display of fluorescent cytoplasmic microtubules was recorded. The grids were then stained with uranyl acetate and the display of fibrous structures recorded by electron microscopy. Thus the display of cytoplasmic microtubular structures in the light microscope and the electron microscope can be compared within the same cytoskeleton. The results show a direct correspondence of the fluorescent fibers in the light microscope with uninterrupted fibers of diameter approximately 550 A in the electron microscope. This is the diameter reported for a single microtubule decorated around its circumference by two layers of antibody molecules. Thus under optimal conditions immunofluorescence microscopy can visualize individual microtubules.