Noguchi M, Furuya S, Takeuchi T, Hirohashi S
Pathology Division, National Cancer Center Research Institute, Tokyo, Japan.
Pathol Int. 1997 Oct;47(10):685-91. doi: 10.1111/j.1440-1827.1997.tb04442.x.
Several simplified fixation methods were examined to determine their suitability for both molecular biological analyses and morphological study. Fixation with 10% v/v formalin alone at 4 degrees C and containing 5 mmol/L ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA) at room temperature preserved significantly more high-molecular-weight DNA than 10% v/v formalin fixation at room temperature. The morphological differences between tissues fixed using these modified formalin fixation methods and conventional 10% v/v formalin fixation were negligible. Of the dehydration fixatives tested, 100% methanol did not cause regional differences due to artificial tissue shrinkage and the morphology of sections prepared by methanol fixation was preserved consistently better than that of acetone- or ethanol-fixed sections. All three dehydration fixatives preserved relatively higher-molecular-weight DNA and RNA, compared with formalin. Cold formalin, formalin containing EDTA at room temperature and 100% methanol are recommended as standard and additional fixatives routine clinicopathological laboratory use.
研究了几种简化固定方法,以确定它们对分子生物学分析和形态学研究的适用性。在4℃单独用10%(v/v)福尔马林并在室温下含有5 mmol/L乙二胺-N,N,N',N'-四乙酸(EDTA)进行固定,与在室温下用10%(v/v)福尔马林固定相比,能显著保留更多的高分子量DNA。使用这些改良福尔马林固定方法固定的组织与传统10%(v/v)福尔马林固定的组织之间的形态差异可忽略不计。在所测试的脱水固定剂中,100%甲醇不会因人为组织收缩而导致区域差异,并且甲醇固定制备的切片形态始终比丙酮或乙醇固定的切片保存得更好。与福尔马林相比,所有三种脱水固定剂都能保留相对较高分子量的DNA和RNA。推荐将冷福尔马林、室温下含EDTA的福尔马林和100%甲醇作为标准固定剂和额外固定剂用于常规临床病理实验室。