Multi-Photon endomicroscope for real-time in vivo vertical sectioning

 描述(申请人提供)：这项生物工程研究资助(BRG)的目标是生产一种新型的多光子显微内窥镜仪器，可以快速改变焦深，从而从体内生物组织产生实时垂直横断面图像，并验证仪器在结肠癌靶向多肽标志物成像方面的性能。与以前的微型多光子仪器相比，建议的仪器将具有更快的轴向扫描速度，以进行无运动伪影(帧速率5-10赫兹)的横断面成像，同时保持较小的直径(3.4毫米)，以便与胃肠道中的小动物成像兼容。仪器性能预计将达到亚细胞分辨率和大视场深组织穿透的目标。仪器的开发将基于驱动、仪器设计、光学和癌症生物学方面的医学和工程专业知识的多学科合作。轴向(组织内)扫描能力将由高速、大行程薄膜PZT垂直平移致动器提供。光纤耦合的远程扫描架构将允许在小型手持仪器中模块化插入优化的光学元件和执行器。仪器验证将通过对标准化目标和/或模体组织的分辨率、穿透深度、帧速率和信噪比进行基准测试来执行。全仪器验证将在基因工程小鼠癌症模型的体内成像研究中进行。小鼠将被标记有靶向的、染料标记的多肽和普通的组织染料。多肽将针对在人类疾病和小鼠模型中展示的特定靶点进行优化。优化后的多肽将被表征为结合亲和力。用不同的荧光试剂对多肽进行活体成像将被用来确定深度多光子成像的最佳操作参数，并表征仪器的性能。

 DESCRIPTION (provided by applicant): The goal of this Bioengineering Research Grant (BRG) is to produce a novel multi-photon microendoscopy instrument that can rapidly vary focal depth and thus produce real-time vertical cross-sectional images from biological tissue in vivo, and to validate instrument performance in imaging of targeted peptide markers for colon cancer. Relative to prior miniature multi-photon instruments, the proposed instrument will feature much faster axial scanning for cross-sectional imaging without motion artifacts (frame rate 5-10 Hz) while maintaining a small diameter (3.4 mm) for compatibility with small animal imaging in the gastrointestinal tract. Instrument performance is expected to meet goals of sub-cellular resolution and deep tissue penetration with substantial field of view. Instrument development will be based on a multi-disciplinary collaboration of medical and engineering expertise in actuation, instrument design, optics, and cancer biology. Axial (into-tissue) scanning capabilities will be provided by high-speed, large- stroke thin-film PZT vertical translational actuators. A fiber-coupled, remote scanning architecture will permit modular insertion of optimized optics and actuators in a small handheld instrument. Instrument verification will be performed through benchmarking against of resolution, penetration depth, frame rate, and signal-to-noise ratio on standardized targets and/or phantom tissues. Full instrument validation will be performed in studies of in vivo imaging of genetically-engineered mouse models of cancer. Mice will be labeled with both targeted, dye-labeled peptides and general tissue dyes. Peptides will be optimized for bonding to specific targets exhibited in human disease and the mouse model. The optimized peptide will be characterized for binding affinity. In vivo imaging of the peptide with different fluorescent reporters will be used to identify optimal operating parameters for deep multi-photon imaging and characterize instrument performance.