Practical high resolution microscopy of uncut, unembedded kidney biopsies

项目摘要 Replikate Technologies开发了多光子显微镜（CHiMP）清除组织学， 提供来自完整病理组织样本的直接数字化图像的处理和成像协议， 例如活组织检查，而不需要蜡包埋、切片或扫描载玻片。适用本 肾脏诊断技术有望大大提高我们提供准确，成本效益高， 并及时诊断患有各种肾脏问题的患者。这也将扩大我们的能力 通过提供其他无法获得的数据来研究和了解肾脏疾病， 新兴的计算机分析工具。 这个直接进入第二阶段的SBIR提案的目的是修改CHiMP的硬件和软件组件 用于肾活检，并验证CHiMP在肾脏诊断方面的非劣效性， 标准协议及其在临床环境中与肾脏病理学工作流程的整合。 CHiMP对于一般组织学具有许多优点，包括：病理学家熟悉的H & E样图像，但 上级图像质量，亚微米有效厚度，无载玻片制备中常见的伪影; 保留完整组织用于辅助分子分析和免疫组织化学的破坏性成像;快速 周转时间，图像在收到组织后几小时内准备就绪;大大降低了劳动力成本;真正的3D 前瞻性;以及用于远程咨询和机器视觉的数字化图像。NIDDK已确定可访问 人类肾脏活检是急性肾功能衰竭精准医学发展的重要瓶颈 和慢性肾病。CHiMP通过保存组织和增加DNA产量直接解决了这一问题。在 此外，该提案将扩展CHiMP的成像，以包括来自胶原的二次谐波产生， 从而能够评估和定量纤维化，纤维化是肾健康的关键因素，并适用于其他疾病。 如肝病。 该提案的具体目标是：1）修改CHiMP显微镜和组织盒，以实现第二次 肾活检中胶原的谐波生成成像，2）在临床环境中收集肾活检标本 并评估与临床工作流程的集成，以及3）添加肾脏特定可视化功能， Stackstreamer，我们的3D可视化软件，并展示了CHiMP诊断的实用性。 成功完成这些目标将为第三阶段过渡到监管批准奠定基础，并提供 这是一个经过验证的平台，可在选定的医院尽早实施。

Project Summary Applikate Technologies has developed Clearing Histology with Multiphoton Microscopy (CHiMP), a novel tissue processing and imaging protocol that provides direct-to-digital images from intact pathology tissue specimens, such as biopsies, without the need for wax embedding, slicing, or scanning of slides. The application of this technology to renal diagnosis is expected to improve considerably our ability to render accurate, cost-effective, and timely diagnoses in patients who suffer from a broad range of kidney problems. It will also expand our ability to study and understand kidney disorders by providing otherwise inaccessible data which can be analyzed with emerging computer analysis tools. The aim of this direct-to-Phase II SBIR proposal is to modify the hardware and software components of CHiMP for application to kidney biopsies and to validate CHiMP's non-inferiority for renal diagnosis compared to standard protocols and its integration into the renal pathology workflow in a clinical setting. CHiMP has many advantages for general histology, including: H&E-like images familiar to pathologists but with superior image quality, sub-micron effective thickness, and without artifacts common to slide preparation; non- destructive imaging that preserves intact tissue for ancillary molecular analysis and immunohistochemistry; rapid turnaround, with images ready within a few hours of receiving tissue; greatly reduced labor costs; true 3D prospective; and digitized images for remote consultation and machine vision. NIDDK has identified access to human kidney biopsies as an important bottle neck for progress in development of precision medicine for acute and chronic kidney disease. CHiMP directly addresses this by preserving tissue and increasing DNA yields. In addition, this proposal will extend CHiMP's imaging to include second harmonic generation from collagen, thereby enabling evaluation and quantitation of fibrosis, a key factor in renal health and applicable to other conditions such as liver disease. The specific aims of this proposal are to: 1) modify the CHiMP microscope and tissue cassettes to enable second harmonic generation imaging of collagen in kidney biopsies, 2) collect renal biopsy specimens in a clinical setting and evaluate integration into the clinical workflow and 3) add renal specific visualization features to Stackstreamer, our 3D visualization software, and demonstrate the utility of CHiMP for diagnosis. 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 in select hospitals.