TECHNICAL SUPPORT CORE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 组织学和显微镜核心设施 组织学和显微镜核心实验室是COBRE的宝贵资产。 然而，这些核心设施受到上一个项目年度发生的严重洪水的严重影响。活动的重点是培训，服务，潜在的应用和协作仪器采购。 大部分设备在洪水期间（10月至3月）无法使用，一些设备在停电期间被用于运行发电机的柴油燃料损坏。 1.仪器：将活细胞成像附件添加到共聚焦显微镜中。该仪器与癌症研究中心共同资助。还安装了教学显微镜和自动组织包埋站，并添加了Leica Tp 1020-自动组织处理器，ArtixScan 400 tf扫描仪和ScanWizard ProTX程序。最后，蔡司荧光显微镜进行了软件程序更新。 2.培训内容：组织学技术员何女士参加了加州组织技术学会年度研讨会（2004年5月13-17日，伯班克，加利福尼亚州）。她接受了基础免疫组织化学新技术方法的培训，并参加了双重免疫染色和质量控制和标准化研讨会。她还在这次会议上接受了激光捕获显微切割方法的培训。 去年举办了一个活细胞成像讲习班，为中心内总共14名研究人员和学生提供培训。两名学生接受了使用LSM 5 Pascal共聚焦显微镜的培训，三名学生接受了Axioskop和两种研究显微镜的指导，两名学生接受了使用Leica旋转/滑动切片机的培训，一名学生接受了Leica振动刀片切片机操作的培训，两名学生使用了Leica石蜡包埋中心，两名学生接受了使用Leica低温恒温器的培训。 DNA测序和基因分型 在与夏威夷癌症研究中心（CRCH）的持续合作伙伴关系中，COBRE与通过CRCH获得的NCI P30赠款的配套资金一起，已用于提供低成本、高通量DNA测序和基因分型设施，该设施利用3100遗传分析仪（应用生物系统公司）。 在2002-03资助年度，共有8，548个测序和基因分型反应，2003-04年度，这一共享设施的使用显著增加。 在过去的一年中，该设施成功地对8，473个样本进行了测序，并对另外5，876个样本进行了基因分型/SNP分析，反应总数为14，349，去年使用量增加了40%。 全基因组表达阵列与生物信息学 在过去的两年里，博伊德博士的企业资金来自联合利华研究公司，心血管研究中心（CVRC）已经成功地建立了一个功能基因组学研究项目，该项目集中于老化的人类面部皮肤中日光性弹性组织变性的病理生物学。 使用来自Affyandum的全基因组表达阵列，该项目确定了几个新的和以前未知的信号通路，这些通路对于理解阳光损伤皮肤过早老化的复杂病理机制至关重要。 对多个DNA阵列分析产生的大数据集进行评估的一个重要先决条件是开发一种小波去噪和奇异值分解算法，以增强对晒伤皮肤内改变的基因表达的检测。 该算法的开发，开发该算法的生物信息学家的招募以及在芯片分析中获得的经验，最近导致了通过国防部成功获得联邦拨款资金，以支持开发用于从全基因组表达阵列获取数据的核心设施，我们新的生物信息学专家的扩大支持，Gordon Okimoto购买了一台新的数据库管理服务器。

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.