Special symposium: fixation and tissue processing models
Special symposium: fixation and tissue processing models
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DOI：
10.1080/10520290903039052
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发表时间：
2009-10-01
影响因子：
1.6
通讯作者：
Grizzle, W. E.
中科院分区：
文献类型：
--
作者：
Grizzle, W. E.
将组织固定在石蜡块上并进行处理，可以切下组织的薄切片（4 - 5 μ m）。组织及其亚细胞成分和周围基质通过切割薄切片并对其进行组织化学或免疫化学染色并使用明视野显微镜观察切片来可视化。在上个世纪，解剖学家和病理学家使用10%中性缓冲福尔马林（10% NBF）作为固定剂的选择。此外，人类和兽医病理学家都接受过使用10% NBF固定的培训，因此这些专业人员不愿意通过使用不同的固定剂来改变诊断组织熟悉的显微镜外观。此外，在大多数研究中，组织加工油对组织微观外观的影响基本上被忽略了。病理组织的石蜡块档案基本上包含10%NBF固定的石蜡块，因此，如果回顾性研究使用档案石蜡块将疾病的分子特征与其结局相关联，则研究必须基于10%NBF固定的油组织。关于10%NBF中的固定如何与组织化学和免疫组织化学染色相互作用的研究数量有限，大多数基于相对较长的固定时间（>= 36 h）。目前，10%NBF中的固定时间已减少至<24 h。关于10% NBF中的固定及其在任何固定时间段内（尤其是短时间内）与组织处理的相互作用知之甚少。关于用10% NBF固定组织如何与使用免疫组织化学、真实的时间定量聚合链反应（PCR）和依赖于从石蜡块中提取的蛋白质分析的技术（包括多重免疫测定或质谱法）的更现代的测定相互作用，知之甚少。一般来说，据报道多种抗体-抗原组合在100%NBF中固定的组织中不起作用，即，对于诸如Ki 67/MIB、雌激素受体α（ER α）和孕激素受体（PR）的抗体-抗原组合，免疫识别几乎完全丧失，对于Bcl-2，免疫识别部分丧失。已经开发了几种模型来研究组织固定和免疫识别的相互作用，但是大多数认为免疫识别的问题完全由固定引起。此外，在这个特殊的研讨会上讨论的一些模型不能预测固定在10% NBF中的冷冻组织的固定效果，而不是处理成石蜡块。本文简要综述了使用10%NBF结合组织处理作为一个相互关联的过程来研究免疫组织化学鉴定的生物标志物的问题。
Fixation and processing of tissue to paraffin blocks permit thin (4-5 mu m) sections of tissues to be cut. Tissues and their subcellular components and Surrounding stroma are visualized by Cutting thin sections and staining them histochemically or immunohistochemically and viewing the sections using a bright field microscope. During the last century, anatomists and pathologists have used fixation with 10% neutral buffered formalin (10% NBF) as the fixative of choice. Also, both human and veterinary pathologists have trained to use fixation with 10% NBF, so these professionals are reluctant to change the familiar microscopic appearance of diagnostic tissues by using different fixatives. In addition, the effects of tissue processing oil the microscopic appearance Of tissue essentially has been ignored in Most Studies. Archives of paraffin blocks of pathological tissue contain essentially paraffin blocks fixed in 10% NBF Therefore, if retrospective studies use archival paraffin blocks to correlate the molecular features of diseases with their Outcomes, the studies must be based oil tissue fixed in 10% NBF. Studies of how fixation in 10% NBF interacts with histochemical and immunohistochemical staining are limited ill number and most are based on relatively long fixation times (>= 36 h) Currently, fixation times, in 10%, NBF have been reduced to < 24 h. Little is known about fixation in 10%, NBF and its interaction with tissue processing for any period of fixation, especially short times. Less is known about how fixation of tissues with 10% NBF interacts with more modern assays using immunohistochemistry, real time quantitative polymerise chain reaction (PCR), and techniques that depend on analysis of proteins extracted from paraffin blocks including multiplex immunoassays or mass spectrometry. In general, multiple antibody-antigen combinations are reported not to work in tissues fixed in 100% NBF, i.e., loss of immunorecognition is nearly complete for such antibody-antigen combinations as Ki67/MIB, estrogen receptor alpha (ER alpha) and Progesterone receptor (PR), and partial for Bcl-2 Several models have been developed to Study the interactions Of tissue fixation and immunorecognition, but most have viewed the problem with immunorecognition as completely caused by fixation. Also, some of the models discussed in this special symposium do not predict the effects of fixation on frozen tissues fixed in 10% NBF and not processed to paraffin blocks. This article is a brief review of issues attending the use of 10% NBF combined with tissue processing as an interrelated process to study biomarkers identified by immunohistochemistry.