Locke M
Department of Zoology, University of Western Ontario, London, Canada.
Microsc Res Tech. 1994 Sep 1;29(1):1-10. doi: 10.1002/jemt.1070290102.
Conventional treatment of tissues for sectioning and transmission electron microscopy uses aldehyde fixation and osmium tetroxide postfixation. Although the result is aesthetically pleasing, osmication destroys some cell components and reduces the chemical activity of others, such as reaction with antibodies and lectins. We have found that aldehyde fixation followed by uranyl acetate preserves and contrasts most structures and visualizes some that are not easily seen after osmication. Aldehyde/UA treated tissues have enough contrast to be observed without section staining while retaining some of the chemical activity that is lost through osmication. Sections of tissues with good preservation and contrast can be used for immunogold and lectin-gold labelling of at least some molecules.
用于切片和透射电子显微镜观察的组织常规处理方法是先用醛类固定,再用四氧化锇进行后固定。尽管结果在外观上令人满意,但锇化处理会破坏一些细胞成分,并降低其他成分的化学活性,如与抗体和凝集素的反应。我们发现,先用醛类固定,再用醋酸铀处理,能保留并使大多数结构形成反差,还能显示出一些在锇化处理后不易看到的结构。经醛类/醋酸铀处理的组织有足够的反差,无需切片染色即可观察,同时还保留了一些因锇化处理而丧失的化学活性。保存良好且有反差的组织切片可用于至少某些分子的免疫金和凝集素金标记。