Practical high resolution microscopy of un-cut, un-embedded lung biopsies

项目摘要和相关性 Applikate Technologies，LLC的这项第二阶段SBIR提案的目的是在成功的 完成我们的第一阶段里程碑，设计、建造和测试原型设备和 用于由实验室人员在临床环境中实施ClearView组织学系统的软件。 Clearview是一种对组织样本进行完整和非破坏性成像的新方法 肺癌和其他癌症。 肺癌活检诊断的错误率可能高达15%。尽管有很多潜力 误差的来源、重要方面与目前组织处理和分析方法的局限性有关。 标准方法限制了诊断病理学家可以看到的组织数量，也就是 可用于个性化药物测试。他们也不愿意咨询非现场专家。 并依赖于受过充分培训的组织技术员的人工处理。应用技术 开发了ClearView，这是一种完整的、非破坏性的、快速的组织处理和成像方法 组织学，通过提供完整的组织可视化来改进基于载玻片的组织学的局限性，减少 准备的人工制品，允许改进定量解释，显示额外的增长模式 信息，并消除了大量的人工准备。ClearView系统生成的图像是 许多情况下，甚至优于传统处理的组织学切片，其伪彩色几乎是 与传统染色无法区分，全部采用完全数字格式，可供远程访问 储藏室。而且ClearView的速度足够快，可以在同一天的两个小时内提供诊断质量的图像 诊断。在我们的第一阶段资助期间，我们还展示了ClearView过程产生了以下订单 更多的DNA用于分子分析，并与免疫组织化学染色兼容。 此外，该方法不仅适用于肺癌的诊断，而且还适用于实际的评估。 任何实体组织癌。 该提议的具体目标是：1)开发一种特定于应用的高速多光子 显微镜及相关软件；2)开发ClearView组织腔原型；3)开发 Clearview原型组织处理器。这三种元素，加上专有试剂，构成了 ClearView系统。在内部测试之后，两个完整的原型系统将被放置在两个 医院，一家是大型三级护理中心，另一家是中型医院，用于现场测试和反馈 由非专家用户在临床环境中使用。这些目标的成功实现将为第三阶段奠定基础 过渡到监管审批，并通过服务工作为早期实施提供经过验证的平台 供学术研究和临床研究团体使用。

Project Summary and Relevance The aim of this Phase II SBIR proposal from Applikate Technologies, LLC, is to build on the successful completion of our Phase I milestones with the design, construction, and testing of prototype devices and software for implementing the ClearView tissue histology system in clinical settings by laboratory personnel. ClearView is a novel approach to performing complete and non-destructive imaging of tissue specimens for lung and other cancers. Lung cancer biopsy diagnosis error rates may run as high as 15%. Although there are many potential sources of error, important aspects relate to limitations of current methods for tissue processing and analysis. Standard methods limit the amount of tissue that can be visualized by the diagnostic pathologist and that is available for personalized-medicine testing. They also are not amenable to consultation with off-site experts and are dependent on manual processing by adequately trained histotechnologists. Applikate Technologies has developed ClearView, a complete, non-destructive, fast, tissue processing and imaging approach to histology that improves on limits of slide-based histology by providing complete tissue visualization, reducing artifacts of preparation, permitting improved quantitative interpretation, showing additional growth pattern information, and eliminating extensive manual preparation. The ClearView system yields images that are, in many cases, superior even to traditionally processed histology slides with pseudo-coloring that is virtually indistinguishable from conventional staining, all in an entirely digital format ready for remote access and storage. And ClearView is fast enough to provide diagnostic quality images in under two hours for same day diagnosis. We have also demonstrated, during our Phase I funding, that the ClearView process yields orders of magnitude more DNA for molecular analyses and is compatible with immunohistochemical staining. Furthermore, the methodology is applicable not only to lung cancer diagnosis, but to evaluation of practically any solid tissue cancer. The specific aims of the proposal are to: 1) Develop an application-specific, high-speed multiphoton microscope and associated software; 2) Develop the ClearView prototype tissue chamber, and 3) Develop the ClearView prototype tissue processor. These three elements, along with proprietary reagents, form the core of the ClearView system. Following in-house testing, two complete prototype systems will be placed in two hospitals, one a large tertiary care center and the second a mid-sized hospital, for on-site testing and feedback by non-expert users in a clinical setting. Successful completion of these aims will set the stage for a Phase III transition to regulatory approval and provide a proven platform for early implementation through service work for academic research and clinical research groups.