Improving biospecimen quality by verifying adequacy at the point-of-acquisition with ex vivo structured illumination microscopy

通过使用离体结构照明显微镜验证采集点的充分性来提高生物样本质量

• 批准号: 8929898
• 负责人: JONATHAN QUINCY BROWN
• 金额: $ 34.08万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8929898
• 关键词: Address; Adopted; Adoption; Advanced Development; Area; Beds; Benign; Biopsy; Biopsy Specimen; Cancerous; Clinical; Collection; Color; Computers; Consultations; Core Biopsy; Development; Diagnosis; Diagnostic; Dyes; Equilibrium; Evaluation; Excision; Fluorescent Dyes; Frozen Sections; Goals; Head; Hematoxylin and Eosin Staining Method; Histopathology; Image; Image Analysis; Internet; Intervention; Left; Lighting; Malignant Neoplasms; Malignant neoplasm of prostate; Methods; Microscopic; Microscopy; Mind; Molecular Analysis; Online Systems; Optics; Pathologist; Pathology; Plague; Procedures; Process; Prostate; Prostatectomy; Publications; Research; Resolution; Sampling; Secure; Sensitivity and Specificity; Site; Slide; Solutions; Specimen; Speed; Staining method; Stains; Structure; System; Technology; Telepathology; Testing; Time; Tissue Banking; Tissue Banks; Tissues; Training; Validation; Work; analog; base; biobank; blind; cancer diagnosis; cancer imaging; cohort; contrast imaging; digital; genetic analysis; imaging modality; improved; in vivo; instrumentation; novel; point of care; prevent; prostate biopsy; public health relevance; sample collection; screening; success; technology development; tumor; virtual; web-enabled

 DESCRIPTION (provided by applicant): Ex vivo microscopy is an emerging set of imaging modalities intended to enable histological imaging of whole tissue specimens at the point-of-procedure. Ex vivo microscopy holds much promise for enabling rapid verification of cancer biospecimen quantity and quality at the time of specimen acquisition. Two areas in which such technologies would be extremely helpful to pathologists are in 1) screening of biopsies collected during a biopsy procedure with the aim of providing guidance for collection of additional tissues, and 2) non-destructively assessing and verifying cancer content in biospecimens collected for banking and downstream molecular analysis. In prostate cancer diagnosis, for example, 70-80% of the 1 million biopsies collected for diagnosis each year are ultimately found to be benign on permanent histopathology review. For tissue banking, the major limiting pre- analytical factor for downstream molecular and genetic analysis is the amount of cancerous tissue present in the banked biopsies. However, currently available in-procedure pathology methods to assess biopsy cancer content (such as frozen section analysis) are too destructive, slow, expensive, and labor-intensive to be widely adopted. Thus, there is a missing opportunity to improve the efficiency and efficacy of diagnostic biopsy, and to guide sample collection and improve cancer biospecimen quality in tissue banking, by non-destructive histological verification. Until now, methods for ex vivo microscopy with the resolution and contrast needed for accurate biopsy verification have been too slow and expensive to realistically be adopted for clinical workflows involving multiple biopsies (16-20 in the case of prostate diagnostic biopsy). Further, the contrast methods (typically topical fluorescent staining with a single dye) have left much to be desired in terms of providing images that leverage the extensive training of pathologists. We have recently developed a set of technologies to address these limitations in an effort to ease adoption. The first is a rapid microscopic optical sectioning scanner, based on video-rate structured illumination microscopy that provides high-resolution, high-contrast images of fluorescently-stained biopsies in seconds at the point-of-procedure. This technology has been optimized for ease-of-use in the pathology workflow, including novel autofocus functions that enable fully automated imaging with minimal user intervention. The second is our recent development of a dual-color fluorescent staining procedure that exactly recapitulates the H&E staining used in standard histopathology. In this R33 Advanced Development project, we will build on these developments to validate VR-SIM as a practical, adoptable ex vivo microscopy solution for point-of-procedure biospecimen verification. In this project we will develop a dual-color version of the VR-SIM system optimized for our fluorescent H&E analog stain, optimize the system for balancing image resolution, contrast, and image acquisition speed, and develop a robust secure web-based multi-resolution image viewer for fast and facile telepathology of the VR-SIM images from any computer with a web browser. The project will culminate in a direct validation challenge of VR-SIM versus frozen section analysis for point-of-procedure biopsy verification in 200 fresh prostate core biopsies.

 描述（由申请人提供）：离体显微镜是一组新兴的成像方式，旨在实现在手术时对整个组织标本进行组织学成像。离体显微镜有望在采集样本时快速验证癌症生物样本的数量和质量。这些技术对病理学家非常有帮助的两个领域是：1）筛选活检过程中收集的活检，目的是为收集其他组织提供指导；2）非破坏性地评估和验证为存储和下游分子分析而收集的生物样本中的癌症内容。例如，在前列腺癌诊断中，每年收集用于诊断的 100 万个活组织检查中，最终通过永久组织病理学检查发现 70-80% 是良性的。对于组织库，下游分子和遗传分析的主要限制性预分析因素是所库活检中存在的癌组织的量。然而，目前用于评估活检癌症内容的术中病理学方法（例如冰冻切片分析）破坏性太大、速度慢、昂贵且劳动密集型，无法广泛采用。因此，缺少通过非破坏性组织学验证来提高诊断活检的效率和功效、指导样本采集和提高组织库中癌症生物样本质量的机会。 到目前为止，具有精确活检验证所需分辨率和对比度的离体显微镜方法太慢且昂贵，无法实际应用于涉及多次活检的临床工作流程（前列腺诊断活检为 16-20）。此外，对比方法（通常是用单一染料进行局部荧光染色）在提供利用病理学家广泛培训的图像方面还有很多不足之处。我们最近开发了一套技术来解决这些限制，以方便采用。第一个是快速显微光学切片扫描仪，基于视频速率结构照明显微镜，可在手术点几秒钟内提供高分辨率、高对比度的荧光染色活检图像。该技术经过优化，易于在病理学工作流程中使用，包括新颖的自动对焦功能，可在最少的用户干预下实现全自动成像。第二个是我们最近开发的双色荧光染色程序，它准确地概括了标准组织病理学中使用的 H&E 染色。在这个 R33 高级开发项目中，我们将在这些开发的基础上验证 VR-SIM 作为一种实用的、可采用的体外显微镜解决方案，用于程序点生物样本验证。在这个项目中，我们将开发一个双色版本的 VR-SIM 系统，针对我们的荧光 H&E 模拟染色进行优化，优化系统以平衡图像分辨率、对比度和图像采集速度，并开发一个强大、安全的基于 Web 的多分辨率图像查看器，以便通过任何具有 Web 浏览器的计算机快速、轻松地对 VR-SIM 图像进行远程病理学。该项目最终将在 200 个新鲜前列腺核心活检中对 VR-SIM 与冰冻切片分析进行直接验证挑战，以进行手术点活检验证。