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Weigners 固定液——一种替代福尔马林固定的组织学固定液,可改善核酸的保存。

Weigners fixative-an alternative to formalin fixation for histology with improved preservation of nucleic acids.

机构信息

Department of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany.

出版信息

Vet Pathol. 2013 Jan;50(1):191-9. doi: 10.1177/0300985812441031. Epub 2012 Apr 26.

Abstract

Formalin fixation and paraffin embedding (FFPE) is the standard method for tissue storage in histopathology. However, FFPE has disadvantages in terms of user health, environment, and nucleic acid integrity. Weigners fixative has been suggested as an alternative for embalming cadavers in human and veterinary anatomy. The present study tested the applicability of Weigners for histology and immunohistochemistry and the preservation of nucleic acids. To this end, a set of organs was fixed for 2 days and up to 6 months in Weigners (WFPE) or formalin. WFPE tissues from the skin, brain, lymphatic tissues, liver, and muscle had good morphologic preservation, comparable to formalin fixation. The quality of kidney and lung samples was inferior to FFPE material due to less accentuated nuclear staining and retention of proteinaceous interstitial fluids. Azan, Turnbull blue, toluidin, and immunohistochemical stainings for CD79a, cytokeratin, vimentin, and von Willebrand factor led to comparable results with both fixates. Of note, immunohistochemical detection of CD3 was possible after 6 months in WFPE but not in FFPE tissues. mRNA, miRNA, and DNA from WFPE tissues had superior quality and allowed for amplification of miRNA, 400-bp-long mRNA, and 1000-bp-long DNA fragments after 6 months of fixation in WFPE. In summary, Weigners fixative is a nonhazardous alternative to formalin, which provides a good morphologic preservation of most organs, a similar sensitivity for protein detection, and a superior preservation of nucleic acids. Weigners may therefore be a promising alternative to cryopreservation and may be embraced by people affected by formalin allergies.

摘要

福尔马林固定和石蜡包埋(FFPE)是组织在组织病理学中储存的标准方法。然而,FFPE 在用户健康、环境和核酸完整性方面存在缺点。Weigners 固定剂已被提议作为人类和兽医解剖学中尸体防腐的替代方法。本研究测试了 Weigners 在组织学和免疫组织化学以及核酸保存方面的适用性。为此,一组器官在 Weigners(WFPE)或福尔马林中固定了 2 天至 6 个月。来自皮肤、大脑、淋巴组织、肝脏和肌肉的 WFPE 组织具有良好的形态保存,与福尔马林固定相当。肾脏和肺组织的质量不如 FFPE 材料,因为核染色不明显,蛋白质间质液保留。阿赞、特恩布尔蓝、甲苯胺蓝和 CD79a、细胞角蛋白、波形蛋白和血管性血友病因子的免疫组织化学染色产生了与两种固定剂相当的结果。值得注意的是,在 WFPE 中固定 6 个月后可以进行 CD3 的免疫组织化学检测,但在 FFPE 组织中不行。来自 WFPE 组织的 mRNA、miRNA 和 DNA 质量较好,在 WFPE 固定 6 个月后,允许扩增 miRNA、400-bp 长的 mRNA 和 1000-bp 长的 DNA 片段。总之,Weigners 固定剂是福尔马林的一种非危险替代品,它为大多数器官提供了良好的形态保存、类似的蛋白质检测灵敏度以及更好的核酸保存。因此,Weigners 可能是冷冻保存的有前途的替代品,并可能被受福尔马林过敏影响的人接受。

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