Cerio R, Dupuy P F, Allen M H, MacDonald D M
J Invest Dermatol. 1986 Oct;87(4):499-503. doi: 10.1111/1523-1747.ep12455571.
The influence of the sequential stages of conventional formaldehyde fixation and paraffin embedding of cutaneous tissue on monoclonal antibody labeling of cell surface antigens is described. The effects of variation in fixation time, dehydration, clearing, wax embedding, and enzyme treatment of cutaneous sections were examined. By curtailing fixation time, using cold ethanol dehydration, and limited cold clearing with xylene, immunoreactivity of several important monoclonal antibodies was retained. Wax embedding could be achieved at 58 degrees C for 1 h or by using low-melting-point wax at 42 degrees C for 3 h. Thus was derived an optimal processing procedure which afforded good tissue morphology and allowed reliable reproducible labeling by monoclonal antibodies to cell surface antigens.
本文描述了皮肤组织常规甲醛固定和石蜡包埋的连续阶段对细胞表面抗原单克隆抗体标记的影响。研究了固定时间、脱水、透明、石蜡包埋以及皮肤切片酶处理等变量的影响。通过缩短固定时间、使用冷乙醇脱水以及用二甲苯进行有限的冷透明处理,保留了几种重要单克隆抗体的免疫反应性。石蜡包埋可在58℃下进行1小时,或使用42℃的低熔点石蜡进行3小时。由此得出了一种最佳处理程序,该程序能提供良好的组织形态,并允许单克隆抗体对细胞表面抗原进行可靠的可重复标记。