1. Preparation, construction and high-throughput automated analysis of human brain tissue microarrays for neurodegenerative disease drug development
- Author
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Richard L.M. Faull, Malvindar K. Singh-Bains, Nasim F. Mehrabi, Adelie Y. S. Tan, and Mike Dragunow
- Subjects
Protocol (science) ,0303 health sciences ,Tissue microarray ,Computer science ,Drug target ,Disease ,In situ hybridization ,Computational biology ,Human brain ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Drug development ,medicine ,Throughput (business) ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
A major challenge in the treatment of neurodegenerative disorders is the translation of effective therapies from the lab to the clinic. One approach to improve this process is the use of human brain tissue microarray (HBTMA) technology to aid in the discovery and validation of drug targets for brain disorders. In this protocol we describe a platform for the production of high-quality HBTMAs that can be used for drug target discovery and validation. We provide examples of the use of this platform and describe detailed protocols for HBTMA design, construction and use for both protein and mRNA detection. This platform requires less tissue and reagents than single-slide approaches, greatly increasing throughput and capacity, enabling samples to be compared in a more consistent way. It takes 4 d to construct a 60 core HBTMA. Immunohistochemistry and in situ hybridization take a further 2 d. Imaging of each HBTMA slide takes 15 min, with subsequent high-content analysis taking 30 min-2 h.
- Published
- 2021
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