Intravital Multiphoton Microscope

描述(由申请人提供)：我们正在为一个多光子激光扫描显微镜系统申请资金。该显微镜系统包括：1)蔡司710NLO多光子成像系统，具有3个光谱探测通道，5条可用于可见光激发共焦成像的激光谱线，用于飞秒脉冲近红外光束；2)带有DIC和高分辨率浸水光学元件的蔡司Axio Imager Z1电动荧光显微镜；以及3)无变色龙相干超快II钛：蓝宝石激光系统。该显微镜将用于检查活的小动物、器官培养物、组织切片、整体器官和已标记了多个参数指示荧光探针的分离组织。双光子和三光子成像将最大限度地减少光损伤和光漂白，最大限度地穿透厚样品，允许用红光和近红外光激发紫外线吸收荧光团，并优化光子收集效率。这一新系统旨在缓解对活细胞和组织进行高分辨率三维无损成像的极高需求。多光子激发显微镜将为我们目前的成像资源提供强大的补充，包括高分辨率磁共振和计算机断层成像。该显微镜还将扩大俄克拉荷马医学研究基金会(OMRF)正在研究的一些转基因小鼠模型的价值。10个主要用户和7个次要用户将立即在OMRF的关节炎和免疫学、心血管生物学、免疫生物学和癌症、自由基和衰老以及细胞、分子和发育生物学等15个独立或合作项目中使用多光子显微镜。预计该仪器将使我们能够获得对几种疾病的病理机制的新见解。OMRF将从这项技术中受益的人类疾病研究的部分清单包括心血管和血栓疾病、炎症和自身免疫、癌症、神经退行性疾病和细菌感染。

DESCRIPTION (provided by applicant): We are requesting funds for a multiphoton laser scanning microscope system. The microscope system consists of: 1) a Zeiss 710 NLO multi-photon imaging system with 3 spectral detection channels, five laser lines for visible light excitation confocal imaging and prepared for femtosecond pulsed NIR beam; 2) a Zeiss Axio Imager Z1 motorized fluorescence microscope with DIC and high resolution water immersion optics; and 3) a Coherent Chameleon free Ultrafast II Ti:Sapphire laser system. The microscope will be used to examine live small animals, organ cultures, tissue slices, whole-mount organs and isolated tissues that have been labeled with multiple parameter-indicating fluorescent probes. Two- and 3-photon imaging will minimize photo-damage and photobleaching, maximize penetration into thick specimens, permit excitation of UV-absorbing fluorophores with red and near infrared light, and optimize photon collection efficiency. The new system is sought to relieve the critically high demand for nondestructive imaging of living cells and tissues with high resolution in 3-dimensions. The multiphoton excitation microscope will provide a strong complement to our current imaging resources, which include high-resolution magnetic resonance and computerized tomography imaging. The microscope will also extend the value of a number of transgenic mouse models being studied at the Oklahoma Medical Research Foundation (OMRF). Ten major users and seven minor users will immediately employ the multiphoton microscope in fifteen independent or collaborative projects in the Programs of Arthritis and Immunology, Cardiovascular Biology, Immunobiology and Cancer, Free Radicals and Aging, and Cell, Molecular & Developmental Biology at OMRF. It is anticipated that the instrument will allow us to access novel insights in the pathological mechanisms of several diseases. A partial list of human disease studies at the OMRF that will benefit from this technology include cardiovascular and thrombotic disorders, inflammation and autoimmunity, cancer, neurodegenerative diseases and bacterial infections.