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在常规透射电镜中,超薄切片的厚度一般不超过0.1微米,否则就看不清样品的细节。但是,使用特殊的标本本浸染方法,选择性地对某些细胞结构染色,就可以在一般透射电镜下观察半薄切片。本实验采用铀—铅—铜浸染技术,选择性地染色一部分细胞器,在75—100KV透射电镜下观察0.25—0.5微米的半薄切片。结果表明,这一技术在超微结构研究中有一定实用意义。实验方法:用2％戊二醛灌注大鼠,取肝、肾、十二指肠和睾丸等组织,切成1立方毫米小块。组织块经缓冲液漂洗后,在42℃下先放入5％醋酸铀水溶液(PH3.5)浸染1小时,再在硝酸铅—硫酸铜—枸椽酸钠溶液中浸染1小时,然后在4℃下用1％饿酸后固定12小时、常规环氧树脂包埋,最后将组织块切成0.25—0.5微米半薄切片,在透射电镜下观察。
In conventional TEM, the thickness of ultrathin sections is usually not more than 0.1 microns, otherwise the details of the sample can not be seen. However, the use of special specimens of this dissemination method, the selective staining of certain cell structures, you can observe the general transmission electron microscopy semi-thin sections. In this experiment, uranium-lead-copper staining technique was used to selectively stain some organelles and observe the semi-thin sections of 0.25-0.5 micrometer under the transmission electron microscope of 75-100KV. The results show that this technique has some practical significance in the study of ultrastructure. Experimental Methods: Rats were perfused with 2% glutaraldehyde, and liver, kidney, duodenum and testis were cut into 1mm3 pieces. Tissue pieces were rinsed with buffer solution and then immersed in 5% aqueous solution of uranyl acetate (PH3.5) at 42 ° C for 1 hour before immersion in lead nitrate-copper sulfate-sodium citrate solution for 1 hour and then at 4 ℃ fixed with 1% sodium glutarate after 12 hours, conventional epoxy resin embedded, and finally cut into pieces 0.25-0.5 micrometre semi-thin section, observed under a transmission electron microscope.