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无甲醛组织学?

Histology without formalin?

作者信息

Buesa René J

出版信息

Ann Diagn Pathol. 2008 Dec;12(6):387-96. doi: 10.1016/j.anndiagpath.2008.07.004. Epub 2008 Sep 9.

DOI:10.1016/j.anndiagpath.2008.07.004
PMID:18995201
Abstract

Because formalin is toxic, carcinogenic, and a poor preserver of nucleic acids, for more than 20 years, there have been numerous attempts to find a substitute, with as many different alternative fixatives, none totally successful. With a fast penetration, formaldehyde is a slow and reversible fixative that requires 24 to 48 hours to completely bind to tissue; thus, any surgical specimen arriving to the laboratory between 8 AM and 4 PM and processed conventionally for the slides to be ready the following day will be only between 30% and 66% bound and even less fixed when the dehydration starts, resulting in an additional and also incomplete alcoholic fixation. This causes infiltration problems and can affect subsequent tests, especially immunohistochemistry. Formaldehyde fixation is tissue thickness independent between 16 microm and 4 mm but is faster at above room temperature, so the fixation of specimens with less than 24 hours in formalin can be improved if the fixing stations in the conventional tissue processors are set at 40 degrees C. If the safety measures are improved to offer a work environment with a time weighted average level of 0.4 ppm, and the contact with formalin is reduced to a minimum by discouraging its neutralization and limiting the recycling practice to filtering methods, formalin could remain as the routine fixative, with modified methacarn for those specimens requiring nucleic acids studies. This is a preferred solution than having to validate all the standard and special procedures, including those US Food and Drug Administration approved, if formalin is replaced by another fixative without its advantages. To the question posed in the title of this article, the answer is "Yes, it can be done, but that is neither likely nor worth it!"

摘要

由于福尔马林有毒、致癌且对核酸的保存效果不佳,二十多年来,人们多次尝试寻找替代品,出现了许多不同的替代固定剂,但都不完全成功。甲醛具有快速渗透的特性,是一种缓慢且可逆的固定剂,需要24至48小时才能完全与组织结合;因此,任何在上午8点至下午4点之间送达实验室并按常规处理以便次日准备好玻片的手术标本,在脱水开始时,其结合程度仅为30%至66%,甚至固定程度更低,这会导致额外的且不完全的酒精固定。这会引发渗透问题,并可能影响后续检测,尤其是免疫组织化学检测。甲醛固定在组织厚度介于16微米至4毫米之间时与厚度无关,但在高于室温时速度更快,所以如果将传统组织处理仪中的固定站设置为40摄氏度,福尔马林固定不足24小时的标本的固定效果可以得到改善。如果安全措施得到改进,以提供一个时间加权平均水平为0.4 ppm的工作环境,并通过不鼓励中和福尔马林以及将回收做法限制为过滤方法来将与福尔马林的接触降至最低,福尔马林可以作为常规固定剂保留下来,对于那些需要进行核酸研究的标本则使用改良甲醇 Carnoy固定液。相较于在没有福尔马林优点的情况下必须验证所有标准和特殊程序(包括那些美国食品药品监督管理局批准的程序),这是一个更优的解决方案。对于本文标题提出的问题,答案是“是的,可以做到,但既不太可能也不值得!”

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