Development of The Prairie Technologies MPLSM to image cancer models in vivo

描述（由申请人提供）：使用多光子激光扫描显微镜（MPLSM）来改善生理相关动物模型的活体成像，对癌症研究具有特别的希望。MPLSM在活体成像方面有很多优点，包括高分辨率、深度切片和提高组织活力。此外，当与二次谐波、荧光寿命和光谱成像方法相结合时，MPLSM有望利用内在的对比度来源，从而消除对外源标签的需要。目前，市场上还没有商用的MPLSM系统能够很好地胜任这项任务。该研究的总体目标是为Prairie Technologies Ultima IV MPSLM增加新的多模态功能，用于肿瘤进展动物模型的活体显微观察。该仪器将允许肿瘤细胞侵入局部基质的成像，通过二次谐波生成（SHG）分析胶原基质的变化，可视化伴随肿瘤生长和进展的代谢变化，以及体内信号分子的研究，特别是那些可能与肿瘤生长、存活、进展、侵袭或转移相关的信号分子。我们提出以下具体目标，通过这些目标，我们将增强Ultima的能力，以实现这些目标。1. 开发用于SHG信号采集和分析的光学和软件方法为Ultima IV开发一个新的采集系统，该系统将由新的软件和硬件组成，可同时实现强度、光谱和寿命的可视化。3. 开发一个自适应光学系统，以纠正系统像差，使图像采集从更深的活体标本。这些动物研究将为未来成像和可视化方法的发展铺平技术道路，这些方法有可能改善人类疾病的诊断和分期。我们设想，一种用户友好的，交钥匙多光子显微镜的发展将促进病理学家使用这项技术。此外，这些技术在未来可能成为外科手术的辅助手段。通过动物模型成像对肿瘤进展的了解将对未来靶向治疗产生积极影响。公共卫生相关性：改进体内多光子成像方法的发展将有助于识别恶性肿瘤的关键特征。了解动物模型中癌症进展的潜在机制可以为人类治疗的未来目标提供建议。

DESCRIPTION (provided by applicant): The use of multiphoton laser scanning microscopy (MPLSM) to improve intravital imaging in physiologically relevant animal models holds particular promise for cancer studies. MPLSM has many advantages for intravital imaging, including high resolution, deep sectioning, and improved tissue viability. Moreover, MPLSM when coupled with second harmonic, fluorescent lifetime and spectral imaging approaches holds the promise of exploiting intrinsic sources of contrast that obviate the need for exogenous labels. Currently, there are no commercial MPLSM systems on the market that are well suited for this task in vivo. The overall objective of the research aims is to add new multimodality functionality to the Prairie Technologies Ultima IV MPSLM for intravital microscopy in animal models of carcinoma progression. This instrument will permit imaging of tumor cells invading into local stroma, analysis of changes in the collagen stroma by second harmonic generation (SHG), visualization of metabolic changes that accompany tumor growth and progression, and investigation of signaling molecules in vivo, especially those that may be relevant to tumor growth, survival, progression, invasion, or metastasis. We propose the following specific Aims, through which we will augment the capabilities of the Ultima to achieve these goals. 1. Develop optical and software approaches for SHG signal collection and analysis 2. Develop an new acquisition system for the Ultima IV that will consist of new software and hardware for simultaneous intensity, spectral and lifetime visualization. 3. Develop an adaptive optics system for correct system aberrations to enable image acquisition from deeper within living specimens. These animal studies will pave the technology for future development of imaging and visualization approaches that have the potential to improve the diagnosis and staging of human disease. We envision that the development of a user-friendly, turn-key multiphoton microscope will facilitate the use of this technology by pathologists. Moreover, these technologies could be a future adjunct to surgery. The understanding of tumor progression that will result from imaging animal models will have a positive impact on targeting future therapies. PUBLIC HEALTH RELEVANCE: The development of improved methods for in vivo multiphoton imaging will help identify key signatures of malignancy of cancer. Understanding the underlying mechanisms of cancer progression in animal models can suggest future targets for human therapy.