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)改善生理相关动物模型的活体成像，为癌症研究带来了特别的希望。MPLSM在活体成像方面具有许多优点，包括高分辨率、深切片和提高组织存活率。此外，当MPLSM与二次谐波、荧光寿命和光谱成像方法相结合时，有望利用内在的对比度来源，从而消除对外部标记的需要。目前，市场上还没有商业化的MPLSM系统能够很好地适应体内的这一任务。这项研究的总体目标是为Prairie Technologies Ultima IV MPSLM增加新的多模式功能，用于癌症进展动物模型的活体显微镜。该仪器将允许对肿瘤细胞侵入局部间质进行成像，通过二次谐波产生(SHG)分析胶原间质的变化，可视化伴随肿瘤生长和进展的代谢变化，以及研究体内的信号分子，特别是那些可能与肿瘤生长、生存、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，我们将增强终极组织的能力，以实现这些目标。1.开发用于倍频信号采集和分析的光学和软件方法2.开发一种新的Ultima IV采集系统，该系统将由新的软件和硬件组成，用于同时显示强度、光谱和寿命。3.开发一种自适应光学系统，用于校正系统像差，以便能够从活体标本内部更深的部位获取图像。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，这些方法有可能改善人类疾病的诊断和分期。我们设想，用户友好的、交钥匙多光子显微镜的开发将促进病理学家使用这项技术。此外，这些技术可能会成为未来外科手术的补充。通过成像动物模型了解肿瘤的进展将对未来的靶向治疗产生积极的影响。公共卫生相关性：体内多光子成像改进方法的开发将有助于识别癌症恶性的关键标志。在动物模型中了解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.