Yamamoto A, Tashiro Y
Department of Physiology, Kansai Medical University, Moriguchi, Osaka, Japan.
J Histochem Cytochem. 1994 Nov;42(11):1463-70. doi: 10.1177/42.11.7930528.
We visualized the surface of ultra-thin sections of rat liver and kidney embedded in LR White resin using an atomic force microscope. The section surface always showed depressions corresponding to the embedded cells at the plasma membrane, at the basal lamina, at mitochondria, and chromatin blocks in the nucleus. The depth of the depression corresponding to the plasma membrane was about 6 nm in both hepatocytes and several types of kidney cells. At the basal lamina, mitochondria, and the chromatin blocks, sections showed deeper depressions of about 10-30 nm. In addition, cytoplasmic surfaces showed strong relief, 3-4 nm on average. The surfaces of the resin block left after ultrathin sectioning showed protrusions corresponding to cells, mitochondria, and the chromatin blocks. In marked contrast, the surface of epon sections was much smoother than that of LR White sections, without showing any marked depressions and relief except for the chromatin blocks of the nucleus. The relation of the surface morphology of the ultrathin sections to the efficiency of immunolabeling is discussed.
我们使用原子力显微镜观察了包埋在LR White树脂中的大鼠肝脏和肾脏超薄切片的表面。切片表面总是显示出与质膜、基膜、线粒体和细胞核中的染色质块处的包埋细胞相对应的凹陷。在肝细胞和几种类型的肾细胞中,与质膜相对应的凹陷深度约为6纳米。在基膜、线粒体和染色质块处,切片显示出约10 - 30纳米的更深凹陷。此外,细胞质表面显示出强烈的起伏,平均为3 - 4纳米。超薄切片后留下的树脂块表面显示出与细胞、线粒体和染色质块相对应的突起。与之形成鲜明对比的是,环氧树脂切片的表面比LR White切片的表面光滑得多,除了细胞核的染色质块外,没有显示出任何明显的凹陷和起伏。本文讨论了超薄切片的表面形态与免疫标记效率之间的关系。