A In order to advance today's basic and translational research in neuroscience, it is imperative that cell/region specific studies can be conducted. Complexity of brain tissues poses a significant challenge when cell specific information has to be derived. Currently there is no commercially available low cost instrument that would permit complex tissue microdissection at cellular resolution and more importantly collection of live cells from both native brain tissues and cell culture dishes. Furthermore, existing laser assisted microdissection instruments are sophisticated and limited by the specific sample pretreatment protocol generally prohibiting its use on native brain tissues. During Phase I, NeuroInDx developed a method for live cell and tissue acquisition that is based on the use of disposable capillary units (DCU-Live) and short, carefully regulated vacuum impulses. Proof of principle experiments demonstrated that the approach is extremely efficient for the microdissection of native and pretreated brain tissues. The cells collected with CTAS-Live are suited for reculturing. Collected samples yield high quality RNA, DNA, and protein for downstream applications. Phase I of this project resulted in the development of a CTAS-Live prototype that features cellular resolution microdissection capabilities from both tissues and cultures without affecting the viability of collected samples. Phase II of this SBIR project will develop CTAS-Live into two commercial models: a base model featuring semiautomatic functions and capability for both microdissection and live cell collection, and a fully automated version of CTAS-Live capable of fast cell and tissue acquisition from multiple specimen via computer controlled calibration process, target recognition and continuous collection controlled via touch screen interface. An important advantage of the proposed instruments is their low price that permits efficient market entrance and commercialization. The base model is priced under $25,000 and the high end automatic version would be priced in the range of $65,000, demonstrating at least four and two times price advantage, respectively versus existing laser based systems. Both base and automated models of CTAS-Live will be thoroughly tested for its performance with a variety of tissue samples and cell cultures focusing on the viability of the acquired cells and hig quality of the macromolecules (e.g. RNA and protein). The epifluorescent module will be tested for the collection of fluorescently labeled cells from tissues. Microdissection of human postmortem tissues will be optimized and the quality of RNA and protein will be evaluated for genomics and proteomics applications. Therefore, aside from initial testing of CTAS-Live prototypes, several Application Notes will be developed offering "sample-to-result" protocols for the potential customers. Developed and "in-house" tested CTAS-Live instruments will be sent to the beta testing sites to ensure end-user satisfaction and obtain valuable feedback. The result of these studies will be successful commercialization of CTAS-Live in Phase III of this project.
NeuroInDx will develop and commercialize a novel cell and tissue acquisition system (CTAS-Live) capable of microdissection at cellular resolution and collection of live cells from both native brain tissues and cell cultures. Proprietary CTAS-Live technology offers a highly efficient and minimally invasive microdissection method permitting collection of viable cells. Development of a base and advanced fully automated version of CTAS-Live instruments will be performed in this Phase II work. The effort will be made to keep the cost low to ensure instrument's affordability. Both instruments will be thoroughly tested for the collection of live cells and microdissection. Specific end-to-end protocols will be developed for the collection of neural progenitor zones, individual cells for recultivation, fluorescently labeled cells from tissue sections and microdissections for genomics and proteomics applications. Both base and automated CTAS-Live instruments will be subjected to beta testing to obtain feedback from the community and identify potential improvement areas.