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组织微阵列的胶带转移切片会引入非特异性免疫组化染色假象。

Tape transfer sectioning of tissue microarrays introduces nonspecific immunohistochemical staining artifacts.

作者信息

Catchpoole D, Mackie N, McIver S, Chetcuti A, Henwood A, Graf N, Arbuckle S

机构信息

Biospecimens Research Group, and Tumour Bank The Children's Cancer Research Unit, The Children's Hospital at Westmead, Westmead, NSW, Australia.

出版信息

Biotech Histochem. 2011 Dec;86(6):421-8. doi: 10.3109/10520295.2010.527859. Epub 2010 Nov 22.

Abstract

Tissue microarrays place tens to hundreds of formalin fixed, paraffin embedded tissue cores into a paraffin block in a systematic grid pattern that permits their simultaneous evaluation in a single section. The fragmented nature of the tissue cores often makes sectioning of tissue microarrays difficult so that the resulting disks of tissue lose their shape, fracture or fall out of the paraffin section altogether. We have evaluated an alternative sectioning protocol for stabilizing the tissue microarray surface by placing an adhesive tape "window" over the face of the paraffin block prior to sectioning. Once sectioned, the tape/sections are transferred directly onto coated microscope slides, thereby avoiding routine floating of sections on a water bath. After sectioning with either the tape transfer or standard protocols, slides were stained either using hematoxylin and eosin or immunohistochemistry using antibodies to S-100 protein and the tissue specific antigens, keratin (AE1/3) and the leukocyte common antigen CD45. We found that the tape method produced thicker sections that were darker and more densely packed with loss of tissue definition compared to sections prepared using water bath flotation. Quantitative image analysis of immunohistochemical staining demonstrated that the tape method produced a higher incidence of nonspecific staining, which raised the potential for false positive staining.

摘要

组织微阵列将数十至数百个经福尔马林固定、石蜡包埋的组织芯以系统的网格模式置于石蜡块中,从而能够在单个切片中对其进行同时评估。组织芯的碎片化性质常常使组织微阵列的切片变得困难,以至于切出的组织圆盘会变形、断裂或完全从石蜡切片中脱落。我们评估了一种替代切片方案,即在切片前在石蜡块表面放置一条胶带“窗口”,以稳定组织微阵列表面。切片后,将胶带/切片直接转移到包被的显微镜载玻片上,从而避免了切片在水浴中常规漂浮的过程。在用胶带转移法或标准方法切片后,载玻片用苏木精和伊红染色,或使用抗S-100蛋白、组织特异性抗原角蛋白(AE1/3)和白细胞共同抗原CD45的抗体进行免疫组织化学染色。我们发现,与用水浴漂浮法制备的切片相比,胶带法产生的切片更厚,颜色更深,组织堆积更密集,组织清晰度下降。免疫组织化学染色的定量图像分析表明,胶带法产生非特异性染色的发生率更高,这增加了假阳性染色的可能性。

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