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Mechanical Compression of Coverslipped Tissue Sections During Heat-induced Antigen Retrieval Prevents Section Detachment and Preserves Tissue Morphology.
Journal of Histochemistry & Cytochemistry ( IF 3.2 ) Pub Date : 2019-01-29 , DOI: 10.1369/0022155419826940
Andreas H Eckhard 1, 2 , Jennifer T O'Malley 1 , Joseph B Nadol 1, 3 , Joe C Adams 1, 3
Affiliation  

Heat-induced antigen retrieval (HIAR) is routinely employed on aldehyde-fixed tissue sections to enhance the reactivity of antibodies that exhibit weak or no specific interactions with tissue antigens when applied in conventional immunohistochemical protocols. A major drawback of HIAR protocols is, however, the heat-induced detachment of sections from the microscope slide with resultant impaired tissue morphology or loss of the section. We developed a method in which tissue sections mounted on glass slides are temporally coverslipped, and a clamp is used to compress the sections on the microscope slide during HIAR treatment. This "pressurized coverslipping" during HIAR was tested on various formalin-fixed tissues (murine kidneys and temporal bones, human tonsils and temporal bones) that were embedded in paraffin or celloidin. The method reliably kept the sections adherent to the slide, preserved the tissue morphology, and effectively retrieved tissue antigens for improved results in immunohistochemical labeling, even for exceptionally delicate, large, and poorly adhering sections, that is, decalcified human temporal bone sections. In summary, we present a simple method for improved slide adherence and morphological preservation of tissue sections during HIAR treatment that can be combined with all HIAR protocols and that requires only basic lab equipment.

中文翻译:

热诱导抗原提取过程中对盖玻片组织切片的机械压缩可防止切片分离并保留组织形态。

当在常规免疫组织化学方案中应用时,通常在醛固定的组织切片上采用热诱导抗原修复(HIAR),以增强与组织抗原之间表现出微弱或没有特异性相互作用的抗体的反应性。但是,HIAR协议的主要缺点是切片从显微镜载玻片上热诱导分离,导致组织形态受损或切片丢失。我们开发了一种方法,其中将安装在载玻片上的组织切片暂时盖上盖玻片,并在HIAR治疗期间使用夹具将其压缩在显微镜载玻片上。HIAR过程中的这种“加压盖玻片”是在石蜡或赛璐oid中包埋的各种福尔马林固定的组织(鼠肾和颞骨,人扁桃体和颞骨)上进行测试的。该方法可靠地保持切片附着在玻片上,保留组织形态,并有效地回收组织抗原,以提高免疫组织化学标记的结果,即使是非常脆弱,大和粘附性差的切片,即脱钙的人颞骨切片。总而言之,我们提出了一种简单的方法,可在HIAR治疗期间改善玻片的粘附性和组织切片的形态保存,可以与所有HIAR方案结合使用,并且只需要基本的实验室设备。
更新日期:2019-11-01
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