Leica TCSSP2RS two-photon microscope for in vivo imaging

用于活体成像的 Leica TCSP2RS 双光子显微镜

• 批准号: 7050299
• 负责人: NICHOLAS R GASCOIGNE
• 金额: $ 50万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2007-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7050299
• 关键词: cell cell interaction; electromagnetic radiation; fasting; immunology; lasers; molecular /cellular imaging; motivation; tissues

DESCRIPTION (provided by applicant): The recent development of two-photon microscopy has made it possible to see within tissues to depths approaching 0.5mm. This makes it feasible to investigate the movements and interactions of cells in living organs. This technology clearly has an important future in immunology. The Intravital two-photon imaging system requested is the Leica Microsystems TCS SP2 RS (resonance scanning) confocal microscope, with infrared laser. This is the state-of-the-art for fast, deep-tissue imaging of motile cells, which is what is required to image cell-cell interactions in tissues such as the lymphoid organs. The user group have a strong background in imaging techniques. They are interested in a wide range of subjects under the broad definition of immunology. There is a particular interest in T cell activation in response to pathogens, cell-cell interactions, tumor growth, and in differentiation. These processes occur in specialized compartments in lymphoid tissues and are therefore not amenable to analysis in vivo by other microscopic techniques. The Leica TCS SP2 RS instrument is based on an upright microscope to allow easy mounting of bulk tissues or living animals, for intravital microscopy. The instrument's MaiTai laser is software tunable to allow relatively fast wavelength changes (within several seconds). The scanning head with resonant galvanometer is a critical component of the system. Resonant scanning is currently the fastest method, approaching video frame rates. The instrument has internal detectors with adjustable bandpass filters. Software controlled motorized slits are used to pass a range of wavelengths split by the prism. This approach is superior to using interference filters because it allows full freedom in optimization of each channel for given fluorophore and has better pass-through efficiency. In addition, 2-channel external (non-descanned) detectors allow maximum utilization of scattered fluorescence. Overall, this instrument will allow the user group to radically improve their ability to perform experiments on understanding the immune system in development and disease.

描述（由申请人提供）：最近两光子显微镜的发展使在组织内看到接近0.5mm的深度成为可能。这使得研究活器官中细胞的运动和相互作用是可行的。这项技术显然在免疫学方面具有重要的未来。所需的插入式两光子成像系统是Leica Microsystems TCS SP2 RS（谐振扫描）共聚焦显微镜和红外激光。这是动向细胞快速，深部组织成像的最先进，这是图像在组织（例如淋巴机构）中图像细胞 - 细胞相互作用所需的。用户组在成像技术方面具有强大的背景。他们对免疫学的广泛定义对广泛的主题感兴趣。响应病原体，细胞 - 细胞相互作用，肿瘤生长和分化，对T细胞激活特别感兴趣。这些过程发生在淋巴组织中的专门隔室中，因此不适合通过其他微观技术在体内分析。 Leica TCS SP2 RS仪器基于直立的显微镜，可轻松安装散装组织或活体动物，用于浸润显微镜。该仪器的Maitai激光器可调节软件，可允许相对快速的波长变化（几秒钟内）。带有谐振电流计的扫描头是系统的关键组成部分。谐振扫描目前是接近视频框架速率的最快方法。该仪器的内部检测器具有可调节的带通滤波器。软件受控的电动缝隙用于传递一系列由棱镜拆分的波长。这种方法优于使用干扰过滤器，因为它可以完全自由地优化给定荧光团的每个通道，并且具有更好的传递效率。此外，2通道外部（未扫描）探测器允许最大程度地利用散射荧光。总体而言，该工具将使用户组从根本上提高他们在了解发育和疾病中的免疫系统方面执行实验的能力。