Core--Molecular detection

描述：分子检测核心(MDC)旨在满足UAB中未得到满足的需求 神经科学界提供灵敏的自动免疫组织化学(IHC)和原位杂交(ISH)检测。该设施的核心将是一个文塔纳发现1系统，能够完全自动化的IHC和ISH应用。该系统与训练有素的技术支持人员相结合，将为新手和经验丰富的IHC/ISH研究人员提供无与伦比的高通量、可重复性的分子定位结果。Discovery系统是唯一能够进行ISH和多标签定位程序的商用自动检测仪器，而MDC将是UAB第一个具有这些能力的任何类型的设施。MDC将提供由核心主管Kevin A.Roth博士与Perkin-Elmer生命科学公司的科学家合作开发的传统和超灵敏酪胺信号放大加(TSA-Plus)检测程序。Roth博士的实验室最近开发的一种使用TSA和量子点标记相结合的灵敏和光稳定的免疫荧光检测的新方法也将推出。MDC还将提供显色和荧光检测选项，以及执行多标记IHC和ISH程序。对于缺乏经验的用户，MDC人员将协助研究人员确定最佳固定条件、组织处理技术和实验设计，以检测感兴趣的神经系统相关分子。为 作为经验丰富的IHC/ISH调查人员，MDC将提供一致的、高质量的标签样本，并协助开发和实施新的检测程序。争取民主变革运动将维护一个网站，在那里可以访问协议，共享图像，并向其他核心用户提出问题。将为研究人员主办一次年度小型专题讨论会，向其他核心用户介绍他们的成果，并帮助促进合作研究工作。这个核心的总体目标是为NINDS支持的研究人员和其他UAB神经科学研究人员提供最先进的分子检测技术，以便更有效和 高效地实现他们的研究目标。我们预计，MDC将成为连接现有UAB校园设施的重要桥梁。因此，我们将从几个组织处理核心实验室中的任何一个接收标本输入，并制作IHC/ISH标记的幻灯片，可以在几个成像设施中的任何一个进行可视化(标准和共聚焦显微镜设施都很容易获得)。或者，研究人员可以提交在他们自己的实验室或使用与MDC相关的组织处理设备准备的细胞和/或组织切片，用于IHC/ISH标记，然后使用MDC或他们自己的实验室提供的显微镜进行可视化。这种很大程度的实验灵活性将允许每个研究人员“定制”他们的MDC使用，以最大限度地满足他们的个人需求。

DESCRIPTION: The Molecular Detection Core (MDC) is designed to fill an unmet need in the UAB neuroscience community for sensitive, automated immunohistochemistry (IHC) and in situ hybridization (ISH) detection. The centerpiece of the facility will be a Ventana Discovery1 System capable of fully automated IHC and ISH applications. This system, combined with well-trained technical support staff, will produce unparaheled high-throughput, reproducible, molecular localization results for both novice and experienced IHC/ISH investigators. The Discovery System is the only commercially-available automated detection instrument capable of ISH and multi-label localization procedures and the MDC will be the first facility of any type at UAB with these capabilities. The MDC will offer both conventional and ultrasensitive Tyramide Signal Amplification-Plus (TSA -Plus) detection procedures developed by Dr. Kevin A. Roth, core director, in collaboration with scientists at Perkin-Elmer Life Sciences. A novel procedure for sensitive and photostable immunofluorescence detection using combined TSA and quantum dot labeling, recently developed by Dr. Roth's laboratory, will also be available. The MDC will also offer both chromogenic and fluorescent detection options as well as perform multi-labeling IHC and ISH procedures. For inexperienced users, MDC personnel will assist investigators in determining optimal fixation conditions, tissue processing techniques, and experimental designs for detection of nervous system-associated molecules of interest. For experienced IHC/ISH investigators, the MDC will provide consistent, high quality labeled samples and assist in the development and implementation of novel detection procedures. The MDC will maintain a website where protocols can be accessed, images shared, and questions asked of other core users. An annual mini-symposium will be sponsored for investigators to present their results to other core users and to help facilitate collaborative research efforts. The overall goal of this core is to provide NINDS supported investigators, and other UAB neuroscience investigators, with state of the art molecular detection techniques so as to more effectively and efficiently achieve their research objectives. We expect the MDC to be an important bridge between pre-existing UAB campus facilities. Thus, we will receive specimen input from any of several tissue processing core laboratories and produce IHC/ISH labeled slides that can be visualized at any of several imaging facilities (both standard and confocal microscopy facilities are readily available). Alternatively, investigators may submit cell and/or tissue sections prepared in their own laboratory or with MDC-associated tissue processing equipment for IHC/ISH labeling with subsequent visualization performed using microscopes available in the MDC or in their own laboratories. This large degree of experimental flexibility will permit each investigator to "customize" their MDC usage to best meet their individual needs.