Köpf I, Hanson C, Delle U, Verbiené I, Weimarck A
Department of Oncology, Göteborg University, Sweden.
Anticancer Res. 1996 Sep-Oct;16(5A):2533-6.
We here present a simplification of the entire procedure for preparing formalin-fixed, paraffin-embedded tissue to be used for FISH-analysis. The steps for deparaffinisation and disintegration of the tissue to produce intact cell nuclei in a monodispersed suspension are detailed as well as the hybridisation steps. The procedure results in a clear cell suspension with intact cell nuclei and without clumped debris particles. The enzymatic digestion is restricted to the trypsinisation step of the deparaffinisation procedure. No further pretreatments prior to hybridization are performed. This modified technique has been successfully applied to different formalin-fixed, paraffin-embedded materials.
我们在此介绍一种简化的用于荧光原位杂交(FISH)分析的福尔马林固定、石蜡包埋组织的制备全过程。详细阐述了组织脱石蜡和分解以在单分散悬浮液中产生完整细胞核的步骤以及杂交步骤。该方法可得到含有完整细胞核且无碎片颗粒聚集的清亮细胞悬液。酶消化仅限于脱石蜡过程中的胰蛋白酶消化步骤。杂交前不进行进一步预处理。这种改良技术已成功应用于不同的福尔马林固定、石蜡包埋材料。