Thiry M
Laboratoire de Biologie Cellulaire et Tissulaire, Université de Liège, Belgium.
Scanning Microsc Suppl. 1996;10:61-70; discussion 70-1.
We have recently combined immunogold labeling procedures with molecular biology methods to pinpoint the precise locations of nucleic acids in biological material at the ultrastructural level. These new immunocytological approaches involve the incorporation of labeled nucleotides in the nucleic acids present at the surface of ultrathin sections prior to immunogold labeling. The antibodies used recognize a nucleoside analogue (bromodeoxyuridine) or a hapten (biotin) employed to label nucleotides. Examples of high-resolution detection include DNA or RNA present in different substructures of cell nuclei, and in particular, in adenovirus-induced intranuclear regions of HeLa cells. In addition to being highly sensitive and specific, these new methods offer the possibility of studying the spatial distribution of nucleic acids in very well preserved, readily recognizable structures.
我们最近将免疫金标记程序与分子生物学方法相结合，以便在超微结构水平上精确确定生物材料中核酸的位置。这些新的免疫细胞化学方法包括在免疫金标记之前，将标记的核苷酸掺入超薄切片表面存在的核酸中。所使用的抗体识别用于标记核苷酸的核苷类似物（溴脱氧尿苷）或半抗原（生物素）。高分辨率检测的例子包括存在于细胞核不同亚结构中的DNA或RNA，特别是在腺病毒诱导的HeLa细胞核内区域。除了高度灵敏和特异之外，这些新方法还提供了在保存非常完好、易于识别的结构中研究核酸空间分布的可能性。
