Automatic Slide Preparation System for Neuropathology Epilepsy Research

用于神经病理学癫痫研究的自动载玻片制备系统

• 批准号: MR/X011860/1
• 负责人: Maria Thom
• 金额: $ 13.66万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX011860%2F1
• 关键词: Automatic; Slide; Preparation; System; Neuropathology

Immunohistochemsitry is a standard technique to demonstrate proteins in tissue sections and has been widely used in epilepsy research, including surgical resections from patients with focal epilepsy as well as post-mortem studies. Quantitative analysis of the distribution of expressed neuronal and glial markers with immunohistochemistry has been used to explore cellular dysfunction of relevance to epileptogenic processes promoting seizures and importantly the consequence of epilepsy, including neuronal loss, glial and vascular dysfunction. The latter is of relevance to co-morbidities in patients with poorly controlled seizures, particularly memory decline. Furthermore regional brain dysfunction in central automomic centres, as the amygdala that regulates respiratory function, is implicated in sudden and unexpected death in epilepsy (SUDEP). Our current research addresses neuropathological alterations that could underpin the brain network changes observed in epilepsy on neuroimaging, neuropsychometrics, functional and automomic studies that cause memory and cognitive decline and can increase SUDEP risk. The primary advantages of quantitative immunohistochemistry is that it enables cellular localisation and regional distribution in human brain samples and can be correlated with parallel genetic and gene-expression studies in study groups compared to controls. Research at our centre is enabled by the Epilepsy society brain and tissue bank, a research tissue bank of donated surgical and post mortem cases collected over 30 years.Quantitative immunohistochemistry is greatly aided by automated systems that improve efficiency, reliability and tracking of immunostained sections enabling seamless antigen detection, slides scanning and digitised automated image analysis. In modern epilepsy neuropathology research, large study groups are required and from many brain regions to correlate with neuroimaging, genetics, clinical outcome and psychometric data as we have demonstrated in our previous publications. Modern immunostaining machines also have additional capabilities including multichannel labelling which allows simultaneous analysis of up to eight antigens, enabling localisation of a disease biomarker with a specific cell type and its relationship to adjacent cells and vascular structures, thus providing insight into disease mechanisms and progression. The current automated immunohistochemistry machine available for epilepsy research is over 12 years old with reduced capacity and functionality and is costly and inefficient to maintain. Our objective is to replace this system with a modern platform that would enhance immunostaining throughput, quality and capability, to improve delivery in our active funded research projects. Our objectives are to ensure this system is adequately maintained and managed and that is made available to epilepsy and neuropathology research staff as well as UCL students who will be trained and fully supported.

免疫组织化学是证明组织切片中蛋白质的标准技术，已广泛应用于癫痫研究，包括局灶性癫痫患者的手术切除以及死后研究。利用免疫组织化学对表达的神经元和神经胶质标记物的分布进行定量分析，已被用于探索与促进癫痫发作的致痫过程相关的细胞功能障碍，以及癫痫的重要后果，包括神经元丢失、神经胶质和血管功能障碍。后者与癫痫控制不佳的患者的合并症有关，特别是记忆衰退。此外，作为调节呼吸功能的杏仁核的中央自动机中枢的区域性脑功能障碍与癫痫（SUDEP）的突然和意外死亡有关。我们目前的研究涉及神经病理改变，这些改变可能支持癫痫患者在神经影像学、神经心理测量学、功能和自动研究中观察到的大脑网络变化，这些变化导致记忆和认知能力下降，并可能增加SUDEP风险。定量免疫组织化学的主要优点是，它可以在人脑样本中进行细胞定位和区域分布，并且与对照组相比，研究组可以进行平行遗传和基因表达研究。我们中心的研究由癫痫协会的脑组织和组织库提供，这是一个30多年来收集的捐赠手术和尸检病例的研究组织库。定量免疫组织化学在很大程度上得益于自动化系统，该系统提高了免疫染色切片的效率、可靠性和跟踪，从而实现了无缝抗原检测、载玻片扫描和数字化自动图像分析。在现代癫痫神经病理学研究中，正如我们在之前的出版物中所证明的那样，需要来自许多大脑区域的大型研究小组来与神经影像学、遗传学、临床结果和心理测量数据相关联。现代免疫染色机还具有额外的功能，包括多通道标记，允许同时分析多达8种抗原，使疾病生物标志物具有特定细胞类型及其与邻近细胞和血管结构的关系，从而提供对疾病机制和进展的深入了解。目前用于癫痫研究的自动化免疫组织化学机器已经使用了12年以上，容量和功能下降，维护成本高，效率低。我们的目标是用一个现代化的平台来取代这个系统，该平台将提高免疫染色的吞吐量、质量和能力，以改善我们积极资助的研究项目的交付。我们的目标是确保该系统得到充分的维护和管理，并将其提供给癫痫和神经病理学研究人员以及伦敦大学学院的学生，他们将接受培训并得到充分支持。