The fundamental studies on the effect of formalin fixation on DNA

福尔马林固定DNA影响的基础研究

• 批准号: 04670180
• 负责人: SUGIYAMA Taketoshi
• 金额: $ 1.15万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670180/
• 关键词: DNA; formalin; restriction enzyme; PCR; lambda-phage; fixation; preservation; 制限・酵素

The effect of formalin fixation on DNA was studied by analysing the effects on lambda-DNA and DNA in animal and autopsied liver tissues.(1)By over-night fixation, formalin split DNA into smaller pieces especially in acidic and saltless conditions at room temperature, although bufferred formalin (BF) in neutral at 4ﾟC prevented DNA fragmentation. On the other hand, lambda-phage DNA became resistent to various restriction enzymes. This effect was dependent on temperature but not on salt concentration of the formalin solution. Thus, these 2 phenomena was considered to be an independent effect of formalin probably caused by scission of phosphate-ester binding for the former, and by cross-linking between the amino groups of the DNA bases for the latter. Regardless of these effects, PCR analysis was successful with lambda-DNA fixed for 2 weeks in 10% formalin and 4 weeks in BF, and with DNA in liver tissues fixed for 6 months in 10% formalin at room temperature. Thus, fixation in BF at 4ﾟC for a short time was recommended as the best fixation method for preservation of DNA in pathology specimens.

通过分析福尔马林固定对动物及尸体解剖肝脏组织的DNA和lambda-DNA的影响，研究福尔马林固定对DNA的影响。(1)通过过夜固定，福尔马林将DNA分裂成更小的片段，特别是在室温下的酸性和无盐条件下，尽管在4℃下的中性缓冲福尔马林（BF）可以防止DNA碎片化。另一方面，噬菌体DNA对各种限制性内切酶产生抗性。这种效应与温度有关，而与福尔马林溶液的盐浓度无关。因此，这两种现象被认为是福尔马林的独立影响，前者可能是由磷酸酯结合的断裂引起的，后者可能是由DNA碱基的氨基之间的交联引起的。不管这些影响如何，将lambda-DNA在10%福尔马林中固定2周，在BF中固定4周，将肝脏组织中的DNA在10%福尔马林中室温固定6个月，PCR分析都是成功的。因此，推荐在4 -丙基酸下短时间固定BF作为保存病理标本DNA的最佳固定方法。

项目成果

Masahiro Koshiba: "The effect of formalin fixation on DNA and the extraction of high-molecular-weight DNA from fixed and embedded tissues" Pathology,Research and Practice. 189. 66-72 (1993)

Masahiro Koshiba：“福尔马林固定对 DNA 的影响以及从固定和包埋组织中提取高分子量 DNA”病理学、研究和实践。

Shuji Hamazaki: "The effect of formalin on restriction endonuclease digestion of DNA and PCR amplification" Pathology,Research and Practice. 189. 553-557 (1993)

Shuji Hamazaki：“福尔马林对 DNA 限制性内切酶消化和 PCR 扩增的影响”病理学、研究和实践。

杉山武敏: "分子病理学" 文光堂, 590 (1993)

杉山武俊：《分子病理学》文科堂，590（1993）

Masahiro Koshiba: "The effect of formalin fixation on DNA and the extraction of high-molecular-weight DNA from fixed and embedded tissues" Pathology,Research and Practice. 189. 58-65 (1993)

Masahiro Koshiba：“福尔马林固定对 DNA 的影响以及从固定和包埋组织中提取高分子量 DNA”病理学、研究和实践。

Taketoshi Sugiyama: Molecular Pathology. Bunkodo, 1993

杉山武俊：分子病理学。