A Novel Technology to Prepare Human Tissue Microarray

DESCRIPTION (provided by applicant): Tissue microarray (TMA) techniques involve arraying cylindrical tissue cores from different specimens on a single slide. TMA's are widely used to identify mRNA or protein markers in human diseases to provide valuable information for the mechanisms of human pathologies and for clinical diagnostics and therapies. Currently TMA is based on paraffin-embedded array technology, i.e., using a small steel tube to take cylindrical tissue biopsies from paraffin-embedded tissues for transfer into a recipient paraffin block. Although this method allows arraying multiple tissue cores on a TMA slide, it has several shortcomings, including severely degraded RNA, and inconsistent protein quality. It is also a time-consuming and expensive process. We have invented a novel TMA technology consisting of a novel array recipient block and new procedures to make TMAs. The experimental procedure involves fixing tissue samples with a solution we developed followed by coring at room temperature. Tissue cores are arrayed into our specially formulated recipient block before sections are cut using a cryostat microtome. The resulting TMA slides can be immediately used for in situ and immunocytochemistry studies without any further treatment. This process yields higher quality RNA and proteins and is also less expensive, less time consuming, simpler and easier than other protocols. In Phase I we will optimize this new technology to produce high density human TMA slides and a cancer cell array. In phase II of the project, we will expand this technology to generate a kit system to make TMAs in a low throughput laboratory setting. This will allow us to produce high quality TMA slides representing a variety of human diseases. Finally, we will develop an automated TMA system for production and use in high throughput screening. The specific aims of this Phase I feasibility study are: 1). Develop a novel TMA technology to produce high-density TMAs that preserve RNA and protein integrity 2). Optimize the tools, reagents and procedures for producing custom high-quality TMAs 3). Develop of cancer cell line arraying methods based on our TMA technology Successful completion of these goals will provide a novel system allowing researchers the ability to easily generate high quality TMA slides. Furthermore, it will prove the feasibility of this approach and provide a firm foundation for the Phase II development of a high throughput TMA system and high quality of disease related TMA slides for clinic diagnosis, drug screening and scientific researches.