Automatic Slide Preparation System for Neuropathology Epilepsy Research

免疫组化是一种标准的技术，以显示蛋白质的组织切片，并已被广泛用于癫痫研究，包括手术切除患者的局灶性癫痫以及尸检研究。免疫组织化学定量分析表达的神经元和胶质细胞标记物的分布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，重要的是癫痫的后果，包括神经元损失，胶质细胞和血管功能障碍。后者与癫痫控制不良患者的合并症有关，特别是记忆力下降。此外，中枢自动神经系统（如调节呼吸功能的杏仁核）的局部脑功能障碍与癫痫猝死（SUDEP）有关。我们目前的研究涉及神经病理学改变，这些改变可能支持在神经影像学，神经心理测量学，功能和自动组学研究中观察到的癫痫脑网络变化，这些变化导致记忆和认知下降，并可能增加SUDEP风险。定量免疫组织化学的主要优点是，它能够在人脑样品中进行细胞定位和区域分布，并且与对照组相比，可以与研究组中的平行遗传和基因表达研究相关。我们中心的研究由癫痫协会脑和组织库支持，这是一个研究组织库，收集了30多年来捐赠的手术和尸检病例。定量免疫组织化学得到了自动化系统的极大帮助，该系统提高了效率，可靠性和免疫染色切片的跟踪，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，需要来自许多大脑区域的大型研究小组与神经成像，遗传学，临床结果和心理测量数据相关，正如我们在以前的出版物中所证明的那样。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，使特定细胞类型的疾病生物标志物及其与相邻细胞和血管结构的关系定位，从而深入了解疾病机制和进展。目前可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，容量和功能都有所减少，维护成本高且效率低。我们的目标是用一个现代化的平台取代这个系统，提高免疫染色的通量、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，并提供给癫痫和神经病理学研究人员以及将接受培训和全力支持的UCL学生。

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.