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）：双光子显微镜的最新发展使得能够观察组织内部接近0.5毫米的深度。这使得研究活器官中细胞的运动和相互作用成为可能。这项技术显然在免疫学领域有着重要的前景。所需的活体双光子成像系统是带有红外激光的 Leica Microsystems TCS SP2 RS（共振扫描）共焦显微镜。这是对运动细胞进行快速深层组织成像的最先进技术，这是对淋巴器官等组织中细胞间相互作用进行成像所需的技术。用户群体在成像技术方面拥有深厚的背景。他们对免疫学广泛定义下的广泛学科感兴趣。人们对响应病原体、细胞间相互作用、肿瘤生长和分化的 T 细胞激活特别感兴趣。这些过程发生在淋巴组织的专门隔室中，因此不适合通过其他显微技术进行体内分析。 Leica TCS SP2 RS 仪器基于正置显微镜，可以轻松安装大块组织或活体动物，用于活体显微镜检查。该仪器的MaiTai 激光器可通过软件调谐，以允许相对快速的波长变化（几秒内）。带谐振振镜的扫描头是系统的关键部件。共振扫描是目前最快的方法，接近视频帧速率。该仪器具有带可调节带通滤波器的内部检测器。软件控制的电动狭缝用于传递由棱镜分裂的一系列波长。这种方法优于使用干涉滤光片，因为它允许完全自由地优化给定荧光团的每个通道，并且具有更好的通过效率。此外，2 通道外部（非退扫描）检测器可最大限度地利用散射荧光。总体而言，该仪器将使用户组能够从根本上提高他们进行实验的能力，以了解发育和疾病中的免疫系统。