Vassy J, Kraemer M, Chalumeau M T
Arch Anat Microsc Morphol Exp. 1984;73(3):181-203.
The influence of the fixation procedure on the localization of albumin and transferrin in adult rat liver has been carried out using an indirect immunoperoxidase technique at the light and electron microscopic levels. Perfusion and immersion fixations with different concentrations of paraformaldehyde (with or without addition of glutaraldehyde) have been investigated. According to the mode of fixation (perfusion versus immersion) and the concentration of the fixative, the number of albumin and transferrin containing hepatocytes could vary from 10% to 100%, and different labeling patterns could be observed at the electron microscopic level. For the same concentration of fixative, a perfusion fixation induces a less intense labeling than an immersion fixation. Thus similar results are obtained after immersion fixation in 6% paraformaldehyde + 0.25% glutaraldehyde or after perfusion fixation in 4% paraformaldehyde + 0.025% glutaraldehyde. Similar data are noticed after immersion fixation in 4% paraformaldehyde or after perfusion fixation in 1% paraformaldehyde + 0.025% glutaraldehyde. Moreover, perfusion fixation induced a more fine cell structure preservation than immersion fixations and avoided the appearance of zones of fixation.
利用间接免疫过氧化物酶技术,在光学和电子显微镜水平上研究了固定程序对成年大鼠肝脏中白蛋白和转铁蛋白定位的影响。研究了用不同浓度多聚甲醛(添加或不添加戊二醛)进行灌注固定和浸入固定的情况。根据固定方式(灌注与浸入)和固定剂浓度的不同,含白蛋白和转铁蛋白的肝细胞数量可在10%至100%之间变化,并且在电子显微镜水平上可观察到不同的标记模式。对于相同浓度的固定剂,灌注固定比浸入固定诱导的标记强度要低。因此,在6%多聚甲醛+0.25%戊二醛中进行浸入固定或在4%多聚甲醛+0.025%戊二醛中进行灌注固定后可获得相似的结果。在4%多聚甲醛中进行浸入固定或在1%多聚甲醛+0.025%戊二醛中进行灌注固定后也可观察到相似的数据。此外,灌注固定比浸入固定能更好地保存细胞的精细结构,并避免出现固定区域。