应用Steedman's wax 切片法观察植物细胞微管骨架

微管骨架在植物发育过程中起重要作用。由于植物细胞的特殊性，与动物细胞相比植物微管骨架的研究遇到更多的困难。简略地介绍了曾被国内外学者应用的植物微管骨架的各种研究方法及其局限性。Steedman's wax是一种多脂蜡。它熔点低(35~37℃)，具有与石蜡相同的切片性质,能够切成不同厚度的连续切片，适合深埋于器官内部的组织或细胞的免疫细胞化学研究。介绍了应用Steedman's wax 切片法观察植物细胞微管骨架的一般程序和方法以及经过作者检验且切实可行的一些技术改进。

Steedman's Wax Sectioning for Observation of the Microtubule Cytoskeleton in Plant Cells

As one of the major components of the cytoskeleton, the microtubule cytoskeleton plays an important role in plant development. The present paper gives a brief introduction on the techniqueswidely applied in this field and their limitations, and recommends a method, a low melting polyester wax (steedman's wax) as an embedding medium for the investigation of the microtubule cytoskeleton with immuno-cytochemistry. Its low melt point (35-37℃) makes it favorable for keeping the antigenicity. Chlorophyll is extracted in the process of infiltration so that the interference of intense autofluorescence from chloroplasts decreases. Steedman's wax has similar characters with paraffin, being able to cut into ribbon-sections of different thickness, which is beneficial both to antibody penetration because of the breaking of cell wall and to the observation of successive focal planes (in ribbon-sections) with confocal laser scanning microscopy. General procedures of Steedman's wax sectioning and some practical improvements were also discussed in the present paper.