Klaushofer K, von Mayersbach H
J Histochem Cytochem. 1979 Dec;27(12):1582-7. doi: 10.1177/27.12.574882.
The suitability of freeze-substitution in n-butanol and paraffin embedding of tissues for the histochemical demonstration of 5'-nucleotidase was investigated and compared with commonly used preparation techniques, such as fresh frozen sections and cryostate sections of cold formalin and glutaraldehyde-fixed rat liver. The influences of each step of the preparation techniques on the enzyme activity were controlled by a quantitative radiochemical assay. Freeze substitution was revealed to be superior to the commonly used preparation techniques with respect to: 1) high sensitivity and specificity of the histochemical 5'-nucleotidase reaction (this is based on the fact that incubation media with very low lead concentrations (0,6 mM/1) can be used); 2) excellent morphological appearance of the tissues showing cytological details of enzyme localization; 3) unlimited storage of the tissue materials and ease of sectioning.
研究了在正丁醇中进行冷冻置换以及组织石蜡包埋用于组织化学显示5'-核苷酸酶的适用性，并与常用的制备技术进行了比较，如新鲜冷冻切片以及冷甲醛和戊二醛固定的大鼠肝脏的低温切片。通过定量放射化学分析控制制备技术各步骤对酶活性的影响。结果表明，冷冻置换在以下方面优于常用的制备技术：1）组织化学5'-核苷酸酶反应具有高灵敏度和特异性（这基于可使用铅浓度极低（0.6 mM/1）的孵育介质这一事实）；2）组织形态良好，能显示酶定位的细胞学细节；3）组织材料可无限期保存且易于切片。
