Rapid quantitative renal fibrosis evaluation with dual-mode microscopy

摘要 肾脏和其他器官一样，具有从急性损伤中恢复的固有能力；然而，严重或 反复损伤可导致慢性肾脏疾病(CKD)，其后遗症可导致82,000人死亡 每年仅在美国。不管最初受伤的原因是什么，通向- 终末期肾病与纤维化(过度或异常的胶原分布)密切相关，其中之一 肾脏疾病严重程度和预后的最重要决定因素。组织学是 评估，通常使用组织化学染色，如三色和PAS，以突出存在 胶原蛋白和基底膜。然而，这些污渍并不是完全特异的，可以 在技术上具有挑战性，以良好和可重复的方式执行，从而有助于观察者之间的可变性和 伴随而来的是诊断准确率的下降。此外，他们还需要准备额外的幻灯片和 额外的染色程序，从而增加了成本，并可能延长诊断过程。 我们提出了优化、部署和测试一种新型显微镜DUET(双模发射和 透射式显微镜)，这将是一种低成本和非常快速的检测解决方案 以及对胶原和其他大分子的存在和分布的数字表征，直接 来自标准福尔马林固定、石蜡包埋的苏木素和曙红染色的玻片。具体来说，我们将 最终确定仪器本身的硬件和软件组件的设计，验证成像 胶原蛋白和其他成分的标准组织学和免疫组织化学染色的性能， 并在我们合作机构(约翰霍普金斯大学和霍普金斯大学)的科学家的帮助下 水牛城)开发用于分析和量化纤维化的强大工具。二重奏乐器硬件将共享 与JHU合作，以确保这些方法在技术上可在多个地点重复使用。 该应用程序利用了光学、生物医学工程、肾脏三个机构的专业知识 病理学和新的人工智能方法。该项目的目标是开发和验证 Duet，它有望成为一种健壮、廉价和实用的方法，用于快速和准确地 对纤维化的评估，可扩展到其他肾脏病理，甚至跨其他器官系统， 对疾病研究、临床实践和患者结局产生重大积极影响。

Abstract Kidneys, like other organs, have an inherent capacity to recover from acute injury; however, severe or recurrent injury can result in chronic kidney disease (CKD), the sequelae of which result in 82,000 deaths annually in the US alone. Regardless of the etiology of the initial injury, the common final pathway leading to- end stage renal disease is closely connected to fibrosis (excess or aberrant collagen distribution), one of the most important determinants of renal disease severity and prognosis. Histology is the gold standard for evaluation, typically using histochemical stains such as trichrome and PAS that highlight the presence of collagens and basement membrane, respectively. Nevertheless, these stains are not completely specific, can be technically challenging to perform well and reproducibly, and thus contribute to interobserver variability and a concomitant decrease in diagnostic precision. Moreover, they also require the preparation of extra slides and additional staining procedures, and thus increase cost and can prolong the diagnostic process. We propose to optimize, deploy, and test a new kind of microscope, DUET (DUal mode Emission and Transmission microscopy), developed at UC Davis, that will be a low-cost and very rapid solution for detection and digital characterization of the presence and distribution of collagen and other macromolecules, directly from standard formalin-fixed, paraffin-embedded hematoxylin and eosin-stained slides. Specifically, we will finalize the design of the hardware and software components of the instrument itself, validate imaging performance against standard histology and immunohistochemical stains for collagen and other components, and with the assistance of scientists at our partnering institutions (John Hopkins University and University of Buffalo) develop robust tools for analysis and quantitation of fibrosis. DUET instrument hardware will be shared with JHU to ensure that the methods are technically reproducible across multiple sites. The application leverages the expertise across three institutions in optics, biomedical engineering, renal pathology and novel artificial intelligence approaches. The goal of the project is development and validation of DUET, which promises to be a robust, inexpensive, and practical approach for the rapid and accurate evaluation of fibrosis, extensible to other renal pathologies, and indeed across other organs systems, with significant positive impact on disease research, clinical practice, and patient outcomes.