Dutt M K
Folia Histochem Cytochem (Krakow). 1978;16(1):47-50.
Treatment of formalin-fixed mammalian tissues with concentrated or 50% phosphoric acid at 5 degrees C for 20 and 50 min. respectively reveals complete extraction of RNA as judged by methyl green followed by staining with pyronin. This procedure also causes depolymerisation of DNA as indicated by the red staining of the nuclei. Sections treated with concentrated phosphoric acid at 5 degrees C for 30 min. causes disruption of the double helical structure of DNA what results in the depression of the pyronin staining. Similarly treated sections show Feulgen positive nuclei. Treatment of sections in 25 % phosphoric acid at 60 degrees C for 15 min. followed by staining with methyl green and pyronin show red nuclei, nucleoli and the cytoplasm. This indicates that extraction of RNA is only possible in cold and not at elevated temperature.
在5℃下分别用浓磷酸或50%磷酸处理福尔马林固定的哺乳动物组织20分钟和50分钟,用甲基绿判断,随后用派洛宁染色,结果显示RNA被完全提取。如细胞核呈红色染色所示,该过程也会导致DNA解聚。在5℃下用浓磷酸处理切片30分钟会导致DNA双螺旋结构破坏,从而导致派洛宁染色减弱。同样处理的切片显示福尔根阳性细胞核。在60℃下用25%磷酸处理切片15分钟,随后用甲基绿和派洛宁染色,细胞核、核仁和细胞质呈红色。这表明RNA的提取只能在低温下进行,而不能在高温下进行。