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

基本信息

批准号：
9314384
负责人：
JONATHAN QUINCY BROWN
金额：
$ 30.24万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-15 至 2019-07-31
项目状态：
已结题

项目摘要

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（由应用程序提供）：离体显微镜是一组新兴的成像方式，旨在使整个组织样品在过程中的组织学成像。离体显微镜在习得时可以快速验证癌症的生物测量量和质量。此类技术对病理学家非常有帮助的两个领域是在1）筛查活检过程中收集的活检，目的是为收集其他时间安排提供指导，以及2）非损害评估和验证收集的用于银行和下游分子分析的生物质量的癌症含量。例如，在前列腺癌诊断中，每年收集的100万活检中有70-80％最终发现在永久性组织病理学评论中是良性的。对于组织库，用于下游分子和遗传分析的主要限制前分析因子是库存活检中存在的癌组织的量。但是，目前可用于评估活检癌症含量（例如冷冻部分分析）的可用过程内病理学方法过于破坏性，缓慢，昂贵且劳动密集型，无法广泛采用。这是缺少的机会来提高诊断活检的效率和效率，并通过非破坏性的组织学验证指导样本收集和改善组织库中癌症生物测量质量。到目前为止，具有准确的活检验证所需分辨率和对比度的离体显微镜的方法太慢且昂贵，无法在涉及多个活检的临床工作流程中采用实际采用（在前列腺诊断活检的情况下为16-20）。此外，在提供利用病理学家的广泛训练的图像方面，对比方法（通常用单个染料使用局部荧光染色）还有很多不足之处。我们最近开发了一系列技术来解决这些限制，以减轻采用。第一个是基于视频速率结构化照明显微镜的快速显微镜光学切片扫描仪，该显微镜在过程中提供了高分辨率的，高分辨率的荧光染色活检图像。该技术已被优化，以易于使用病理学工作流程，包括新型的自动对焦功能，可通过最少的用户干预实现全自动成像。第二个是我们最近对双色荧光染色程序的开发，该程序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，我们将基于这些开发项目，以验证VR-SIM作为一种实用，适应性的离体显微镜解决方案，以实现处理点生物循环验证。在此项目中，我们将开发针对我们的荧光H＆E模拟不锈钢优化的VR-SIM系统的双色版本，优化了平衡图像分辨率，对比度和图像采集速度的系统，并开发出可靠的基于Web的安全性多分辨率图像查看器，用于从任何计算机中使用计算机的VR-SIM图像的快速和宽敞的图像远处。该项目将在200个新的前列腺核活检中的VR-SIM与冷冻部分分析的直接验证挑战中达到直接验证挑战。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nondestructive Diagnosis of Kidney Cancer on 18-gauge Core Needle Renal Biopsy Using Dual-color Fluorescence Structured Illumination Microscopy.

DOI：
10.1016/j.urology.2016.08.036
发表时间：
2016-12
期刊：
Urology
影响因子：
2.1
作者：
Liu J;Wang M;Tulman D;Mandava SH;Elfer KN;Gabrielson A;Lai W;Abshire C;Sholl AB;Brown JQ;Lee BR
通讯作者：
Lee BR

Persistent Homology for the Quantitative Evaluation of Architectural Features in Prostate Cancer Histology