Core--Molecular detection

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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.