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非交联固定剂 RCL2®-CS100 与病理诊断和分子分析兼容。

The non-crosslinking fixative RCL2®-CS100 is compatible with both pathology diagnosis and molecular analyses.

机构信息

Department of Pathology, Val d'Aurelle Cancer Institute, Montpellier 34 298, France.

出版信息

Pathol Oncol Res. 2013 Jan;19(1):41-53. doi: 10.1007/s12253-012-9556-2. Epub 2012 Aug 15.

Abstract

Formalin is the key agent for tissue fixation and pathological diagnosis. However, it poorly preserves nucleic acids and this can impair molecular studies. An alternative to formalin would be a fixative which can allow both morphologic and molecular analyses. To assess the suitability of such a fixative, breast (n = 11) and colon (n = 12) tumor samples were fixed in the non cross-linking RCL2®-CS100 fixative and compared to paired formalin-fixed and to frozen samples, the current standards for histology and molecular analyses, respectively. Sections from RCL2®-CS100-fixed samples showed good preservation of cellular and architectural morphology, suitable for routine diagnosis. Although some antibodies required change in the immunohistochemical procedures, quality of the immunohistochemical staining was comparable to that obtained after formalin fixation. HER2 chromogenic in situ hybridization was also successfully performed. High quality DNA could be isolated from RCL2®-CS100-fixed cancer tissues as evidenced by successful amplification of large DNA fragment, CGH array, KRAS and microsatellites genotyping. The quality of RNA from RCL2®-CS100-fixed samples was slightly decreased in comparison to that of RNA isolated from frozen samples, as evidenced by a decreased RNA integrity number but remained exploitable for molecular assays. Our results support the use of the RCL2®-CS100 fixative for histological diagnosis and recovery of high-quality nucleic acids for molecular applications. However, specific procedures for tissue handing and processing, essential to provide high-quality specimens, could limit its use to small target lesions which cannot be frozen without impairing their pathological evaluation.

摘要

福尔马林是组织固定和病理诊断的关键试剂。然而,它对核酸的保存效果较差,这可能会影响分子研究。福尔马林的替代试剂应该是一种既能进行形态学分析又能进行分子分析的固定剂。为了评估这种固定剂的适用性,我们分别用非交联的 RCL2®-CS100 固定剂固定了 11 例乳腺和 12 例结肠肿瘤样本,并与配对的福尔马林固定样本和冷冻样本进行了比较,后两者分别是组织学和分子分析的当前标准。RCL2®-CS100 固定样本的切片显示出良好的细胞和组织结构形态保存,适合常规诊断。尽管一些抗体在免疫组化过程中需要改变,但免疫组化染色的质量与福尔马林固定后获得的质量相当。HER2 显色原位杂交也成功进行。从 RCL2®-CS100 固定的癌症组织中可以分离出高质量的 DNA,这可以通过成功扩增大片段 DNA、CGH 阵列、KRAS 和微卫星基因分型来证明。与从冷冻样本中分离的 RNA 相比,RCL2®-CS100 固定样本中的 RNA 质量略有下降,这表现在 RNA 完整性数量降低,但仍可用于分子检测。我们的结果支持使用 RCL2®-CS100 固定剂进行组织学诊断,并回收高质量的核酸用于分子应用。然而,为了提供高质量的标本,组织处理的特定程序可能会限制其在不能冷冻而不影响其病理评估的小靶病变中的应用。

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