ZW10相互作用蛋白(ZWINT)参与线粒主轴检查点和癌的发病机制。在这里,我们介绍了一种在人类肺癌组织中对ZWINT进行免疫染色的方法,然后对整个幻灯片进行数字扫描和图像分析。该方法可以提供高质量的数字图像和可靠的结果。
本研究旨在介绍人体肺组织的免疫染色方法,然后进行全滑数字扫描和图像分析。数字扫描是扫描一叠幻灯片并生成高质量数字图像的快速方法。它可以产生与病理学家的传统光显微镜(CLM)一致的结果。此外,数字图像的可用性使得多人可以同时观察到同一张幻灯片。此外,幻灯片的数字图像可以存储在数据库中,这意味着玻璃幻灯片的长期恶化是可以避免的。此技术的局限性如下。首先,它需要高质量的制备组织和原始的免疫组织化学(IHC)幻灯片,没有任何损伤或过多的密封剂残留物。其次,肿瘤或非肿瘤区域应在使用软件进行分析之前由经验丰富的病理学家指定,以避免在评分过程中对肿瘤或非肿瘤区域产生任何混淆。第三,操作员需要在整个幻灯片成像过程中控制色彩再现。
ZW10相互作用蛋白(ZWINT)是血吸管复合物的必要成分,它涉及线粒主轴检查点1,2,3。据报道,ZWINT的耗竭导致异常的过早染色体分离1,2,3。最近的研究表明,ZWINT通过促进肿瘤细胞4、5的增殖参与多种肿瘤的发病机制。我们之前曾报道过,ZWINT在肺癌5中过度表达。人们普遍认为,病理学家使用CLM分析幻灯片是费时费时的,而不是定量的6,7,8。 此外,存储的玻璃幻灯片的损坏可能使其无法收回以前创建的幻灯片。基于计算机的数字化全幻灯片成像(WSI)的新兴方法可以克服这些限制6,7,8。
为此,我们描述了人类肺癌组织中ZWINT免疫染色的方法,以及全幻灯片数字扫描和基于软件的图像分析。该方法的主要优点是与 CLM 产生一致的结果。该技术可广泛应用于血氧素-eosin染色(H&E)和IHC的病理评分、原位杂交(FISH)、组织微阵列(TMA)以及药物发现和开发等领域。
全幻灯片扫描正成为一个热门话题,其强大的扫描和生产高品质的图像,用于临床和研究目的11,12,13。图像可以在11、12、13分钟之内通过滑动扫描显微镜产生。通过应用该方法,我们获得了ZWINT IHC幻灯片的高质量图像,并比较了肿瘤组织和非肿瘤组织的H-分数。在…
The authors have nothing to disclose.
该项目得到了中国国家自然基金会(第81500151号、81400121号、81270607、81541027和81501352)和湖北省自然基金会(中国)(第2017CFB631号)的支持。作者对郭勤、常敏、李辉及其武汉谷歌生物科技有限公司同事的技术支持表示感谢。作者还感谢穆罕默德·贾迈勒的语言编辑。
Pannoramic MIDI | 3D HISTECH | Cat: PMIDI-040709 | An automated digital slide scanner with a remarkable feature set :12-slide capacity, fluorescence scanning, and many more. |
QuantCenter | 3D HISTECH | Downloaded from the official website of the company | The framework for 3DHISTCH's image analysis applications. |
LEICA RM2235 | Leica Microsystems | Cat: 14050038604 | The enhanced precision of the new accessories will add convenience to block to knife approach as well as specimen orientation. |
Rabbit anti-human Anti-ZWINT antibody | Abcam | Cat: ab197794 | Immunohistochemical analysis of ZWINT in human lung tissue. |
Anti-rabbit secondary antibody | Wuhan Goodbio Technology | Cat:GB23303-1 | Secondary antibody for IHC staining. |
Phosphate-buffered saline | Wuhan Goodbio Technology | Cat:G0002 | A solution containing a phosphate buffer. |
OLYMPUS CX23 | OLYMPUS | Cat:6M87620 | Microscope for detection of H&E or IHC slides. |
Dimethylbenzene | Shanghai Lingfeng Chemical Reagent | Cat:1330-20-7 | A colorless, flammable fluid used as a solvent and clarifying agent in the preparation of tissue sections for microscopic study. |
Hematoxylin Staining Solution | Wuhan Servicebio technology | Cat:G1039 | It is commonly used for histologic studies, oftern colors the nuclei of cells blue. |
Tween 20 | Baitg | Cat:2005-64-5 | It is a polysorbate-type nonionic surfactant formed by the ethoxylation of sorbitan before the addition of lauric acid. It is used as a deterent and emulsifier in pharmacological applications. |
Citric acid repair liquid | Wuhan Servicebio technology | Cat:G1202 | Is is used to repair antigen after fixation during IHC procedure. |
LEICA ASP200s | Leica | Cat: 14048043626 | It was designed for routine and research histopathology of up to 200 cassettes. |
LEICA Arcadia H | Leica | Cat: 14039354103 | It is a heated paraffin embedding station and allows for simple operation and precise control, resulting in improved quality, a smooth workflow and reliability. |
LEICA Arcadia C | Leica | Cat: 14039354102 | It is a cold plate holding more than 60/65 cassettes on its large working surface. It was designed with an environment adaptive control module to make sure the operating temperature is always stabilized at -6°C. |
CaseViewer Software | 3DHISTECH |