Open-top light-sheet microscopy for non-destructive 3D pathology of prostate biopsies

摘要 我们提出了一种新的方法，前列腺癌病理学使用3D开顶光片 （OTLS）显微镜和光学透明。约20%的初始阴性患者 前列腺活检程序在随后的活检中发现有前列腺癌 procedure.以前的研究表明，检查100%的活检组织， 前列腺癌的检出率，但这些研究使用劳动密集型手动切片 这些技术对于临床实验室来说是实用的。我们假设100%的 使用OTLS显微镜检查的前列腺活检组织，而不是使用电流 病理学技术，可以检测到隐匿性癌，否则将错过在一个 患者的子集。此外，我们打算表明，三维OTLS可以无缝集成 临床实验室的工作流程。该提案的具体目标是： 我们的OTLS方法不干扰福尔马林固定的当前病理学实践， 石蜡包埋（FFPE）组织切片，苏木精和伊红（H&E）组织学，以及 免疫组织化学（IHC），以及2）显示3D病理学可以检测隐匿性癌症 我们将比较“伪H &E”3D OTLS图像的诊断 传统的H&E病理学我们将使用存档的FFPE活检组织块， 前列腺癌患者的初始活检结果为阴性。 通过本提案中的研究，我们将验证我们的创新3D 具有标准病理学实践的OTLS方法，这是在 病理学社区和监管考虑。我们还将提供初步的 支持3D病理学临床价值的证据。该项目旨在实现IMAT的目标， 开发大大改进的癌症检测和风险评估技术。

Abstract We are proposing a new approach for prostate cancer pathology using 3D open-top light-sheet (OTLS) microscopy and optical clearing. Approximately 20% of patients with an initial negative prostate biopsy procedure are found to have prostate cancer on a subsequent biopsy procedure. Previous studies have shown that examination of 100% of biopsy tissue improved prostate cancer detection rate, but these studies used labor-intensive manual sectioning techniques that are practical for a clinical laboratory. We hypothesize that imaging 100% of prostate biopsy tissue using OTLS microscopy, rather than 1-2% of the biopsy using current pathology technology, can detect occult carcinoma that would have otherwise been missed in a subset of patients. Furthermore, we intend to show that 3D OTLS can be seamlessly integrated into the current clinical laboratory workflow. The specific aims of this proposal are: 1) Validate that our OTLS methods do not interfere with the current pathology practice of formalin-fixed paraffin-embedded (FFPE) tissue sections, hematoxylin and eosin (H&E) histology, and immunohistochemistry (IHC), and 2) To show that 3D pathology can detect occult cancer missed by traditional microscopy, we will compare diagnosis of “pseudo-H&E” 3D OTLS images to traditional H&E pathology. We will use archived FFPE biopsy blocks from patients with prostate cancer who had a negative initial biopsy result. Through the studies in this proposal, we will validate the non-interference of our innovative 3D OTLS methods with standard pathology practice, a necessary prerequisite for adoption within the pathology community and for regulatory considerations. We will also provide preliminary evidence to support the clinical value of 3D pathology. This project addresses the IMAT goal of developing substantially improved cancer detection and risk assessment technologies.