TECHNICAL SUPPORT CORE

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Histology and microscopy core facility The histology and microscopy core laboratory are invaluable asset to the COBRE. However, these core facilities were substantively impacted by the severe flood that occurred during the past project year. Activities have focused upon training, service, potential applications and collaborative instrumentation acquisition. Much of the equipment was out of service during the flood (Oct-March), some sustained damage from deisel fuel that was used to run generators during the power outage. 1. Instrumentation: A live cell imaging attachment was added to the confocal microscope. This instrument was co-funded with the Cancer Research Center. A teaching microscope and an automated tissue embedding station were also installed and a Leica Tp 1020-Automatic Tissue Processor, ArtixScan 400tf scanner with ScanWizard ProTX programs were added. Finally, the Zeiss flourescent microscope underwent a software program update. 2. Training: Ms He, the histology technician, participated in the Annual Symposium of the California Society for Histotechnology (May 13-17, 2004 in Burbank, CA. She received training in novel technical methods in basic immunohistochemistry and participated in the Double Immunostaining and Quality Control and Standardization workshops. She also received training at this meeting in laser capture micro-dissection methods. A Live Cell Imaging Workshop was ofered in the last year that provided training for a total of 14 investigators and students within the Center. Two students were trained in the use of the LSM 5 Pascal confocal microscope, three students received instruction in the Axioskop and the two research microscopes, two students were trained to use the Leica rotary/sliding microtome, one student was instructed in the operation of the Leica vibrating blade microtome, two students used the Leica paraffin embedding center and two students were trained in the use of the Leica cryostat. DNA Sequencing and Genotyping In a continuing collaborative partnership with the Cancer Research Center of Hawaii (CRCH), the COBRE, with matching funds from an NCI P30 grant through CRCH, has been used to provide a low cost, high throughput DNA sequencing and genotyping facility that utilizes a 3100 Genetic Analyzer (Applied Biosystems). From a total of 8,548 sequencing and genotyping reactions during the 2002-03 grant year, the use of this shared facility has grown significantly in the year 2003-04. During this last year, the facility successfully sequenced 8,473 samples and performed genotyping/SNP analyses on another 5,876 samples for a total number of reactions of 14,349, a 40% increase in use in the last year. Whole genome expression arrays and bioinformatics Over the last two years, with Dr. Boyd's corporate funding from Unilever Research Inc., the Cardiovascular Research Center (CVRC) has successfully established a functional genomics research project focused upon the pathobiology of solar elastosis in aging human facial skin. Using whole genome expression arrays from Affymetrix, this project identified several new and previously unknown signaling pathways critical to the understanding of the complex pathomechanisms of premature aging of sun-damaged skin. A vital prerequisite to the evaluation of the large data sets generated from the analysis of multiple DNA arrays was the development of a wavelet denoising and singular value decomposition algorithm for the enhanced detection of altered gene expression within sun-damaged skin. The development of this algorithm, the recruitment into CVRC of the bioinformatician that developed it and the experience gained in chip analysis has led recently to the successful acquisition of federal appropriation funds, through the Department of Defense, to support the development of a core facility for the acquisition of data from whole genome expression arrays, the expanded support of our new bioinformatics specialist, Gordon Okimoto and the purchase of a new server for database management.