Gowen B E, Buendia B, Karsenti E, Fuller S D
Structural Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Histochem J. 1995 Mar;27(3):240-6.
Accurate ultrastructural localization of the components of centrosomes is an important step toward the determination of their function. We have used an electron microscopy procedure to preserve centrosome-associated antigens which enables their high-resolution localization. The unique part of our procedure is the application of a post-sectioning fixation step which overcomes the poor section contrast and morphological appearance that limits the use of low-temperature processing and Lowicryl embedding. The efficacy of our approach is demonstrated by the efficient labelling of alpha-tubulin in the well-preserved and contrasted microtubule barrels of the centrides of isolated mammalian centrosomes.
准确对中心体各成分进行超微结构定位是确定其功能的重要一步。我们采用了一种电子显微镜方法来保存与中心体相关的抗原，从而能够对其进行高分辨率定位。我们方法的独特之处在于应用了切片后固定步骤，该步骤克服了限制低温处理和Lowicryl包埋使用的切片对比度差和形态外观不佳的问题。通过对分离的哺乳动物中心体中心粒保存良好且对比度高的微管桶中的α-微管蛋白进行有效标记，证明了我们方法的有效性。
