Cancer histology and QC via MUSE: Sample-sparing UV surface-excitation microscopy

通过 MUSE 进行癌症组织学和质量控制：保留样品的紫外表面激发显微镜

• 批准号: 9233713
• 负责人: RICHARD M. LEVENSON
• 金额: $ 37.36万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-08 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9233713
• 关键词: Appearance; Area; Basic Science; Biological; Biological Assay; Biopsy; Biopsy Specimen; Cancer Histology; Cell Survival; Cells; Clinic; Clinical; Color; Computer software; Contrast Media; DNA; Detection; Development; Diagnostic; Disease; Dyes; Effectiveness; Ensure; Eosine Yellowish; Epithelial; Experimental Animal Model; Exposure to; Fluorescence; Fluorescent Dyes; Formalin; Frequencies; Fresh Tissue; Geometry; Goals; Harvest; Histology; Image; Imaging Device; Label; Lasers; Lesion; Manuals; Methods; Microscope; Microscopy; Microtomy; Molecular; Molecular Analysis; Morphologic artifacts; Nervous System Neoplasms; Optical Coherence Tomography; Optics; Organ; Paraffin Embedding; Pathologist; Pathology; Patients; Penetration; Preparation; Process; Proteins; RNA; Research Personnel; Resolution; Sampling; Services; Signal Transduction; Slide; Source; Specimen; Staining method; Stains; Surface; Testing; Time; Tissue Sample; Tissue Viability; Tissues; Toxicology; Translational Research; Ultrasonography; Ultraviolet Rays; Validation; Visible Radiation; Xenograft procedure; base; biobank; cancer diagnosis; cost; file format; high resolution imaging; imaging capabilities; instrumentation; lens; novel; point of care; sample fixation; software development; tissue processing; tool; tumor; validation studies

PROJECT SUMMARY: Pathology—determining the causes and effects of disease, often by using a microscope to examine patients' tissue—is essential for arriving at a correct cancer diagnosis, but it has become increasingly important to submit relevant portions of often tiny tissue samples for DNA and other molecular and functional tests. Making sure that the submitted material actually contains tumor, in sufficient quantity, is not always easy, and sometimes just preparing conventional microscope slides can consume most or even the entire specimen. We have developed a new, simple and inexpensive approach we term MUSE, for Microscopy with UV Surface Excitation, that can provide high-resolution images directly and quickly from fresh tissue without consuming it, and thus can preserve intact, high-quality specimens for biobanking, downstream molecular and functional analyses. MUSE, by ensuring that acquired small biopsy specimens are indeed fit for purpose, can avoid the need to have a patient return at a later date for additional biopsies. In addition, the method will allow for lesion detection over larger cross-sections of tissue, such as organ whole-mounts, not currently practical with conventional instrumentation. Finally, we expect that MUSE will yield additional biologically informative imaging by avoiding artifacts inherent in conventional tissue processing and sectioning. We focus here on facilitation of tissue-based molecular studies, viable tumor harvest for xenografts and cultured spheroids, and biobanking, and include crucial validation studies to ensure that the MUSE process does not compromise sample utility due to possible impacts from exposure to intercalating dyes and/or UV light. MUSE relies on two mechanisms: 1) surface-restricted excitation of fluorescent dyes due to micron- scale penetration of sub-300-nm ultraviolet light; and 2) the fact that many conventional dyes excited in this way emit visible light. These signals are bright enough to be detected by conventional color cameras using sub-second exposure times, allowing rapid imaging of large areas. MUSE eliminates any requirement for conventional histology processing with formalin fixation, paraffin embedding, or thin-sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography instrumentation, can eventually cost in the range of a few thousands of dollars, and therefore affordable at the point of care (point of biopsy). MUSE samples are stained within seconds using familiar histology stains, such as eosin and DAPI, and the resulting high-resolution images converted from fluorescence to H&E-like brightfield appearance for interpretation in real time (but with novel and potentially useful features) easily interpreted by pathologists. While rapid cellular-scale imaging of fresh tissue can have significant benefits in clinical arenas, it can also empower basic and translational research use for essentially instant histology-, pathology- or toxicology-relevant images directly from experimental animal models—at the bench—and may help relieve such investigators from having to rely on often overworked, sometimes unavailable, conventional histology and pathology services.

项目摘要：病理学 - 确定疾病的原因和影响，通常是通过使用 显微镜检查患者的组织 - 对于到达正确的癌症诊断至关重要，但它具有 提交通常微小的组织样本的DNA和其他相关部分变得越来越重要 分子和功能测试。确保提交的材料实际上包含肿瘤，足够 数量，并不总是那么容易，有时只是准备常规显微镜幻灯片可能会消耗大多数 甚至整个标本。我们已经开发了一种新的，简单且廉价的方法，我们称为缪斯女神 带有紫外表面激发的显微镜，可以直接从新鲜提供高分辨率图像 组织不食用它，因此可以保留完整的高质量标本，用于生物群，下游 分子和功能分析。 缪斯，通过确保获得的小型活检标本确实适合目的，可以避免 需要让患者在以后返回其他活检。此外，该方法将允许病变 检测较大的组织的横截面，例如器官的整数，目前不实用 常规仪器。最后，我们预计缪斯将产生更多的生物学内容丰富的成像 通过避免传统组织加工和切片固有的伪影。我们在这里专注于 基于组织的分子研究，用于异种移植物和培养的球类动物的可行肿瘤收获以及生物群， 并包括关键的验证研究，以确保缪斯过程不会损害样本实用程序 暴露于互化染料和/或紫外线的可能影响。 缪斯依赖于两种机制：1）由于微米而引起的荧光染料的表面限制兴奋 低300 nm紫外线的尺度渗透； 2）许多传统染料对此感到兴奋的事实 方式发出可见光。这些信号足够明亮，可以通过使用常规彩色摄像机检测到 次要暴露时间，可以快速成像大面积。 缪斯消除了使用福尔马林固定的传统组织学处理的任何要求，石蜡 嵌入或薄色。它不需要激光，共聚焦，多光子或光学相干断层扫描 仪器有时会花费数千美元的价格，因此在 护理点（活检点）。缪斯样品在几秒钟内使用熟悉的组织学污渍染色，例如 作为曙红和DAPI，以及所得的高分辨率图像从荧光转换为H＆E样图像 Brightfield出现实时解释（但具有新颖且潜在有用的功能）很容易 由病理学家解释。 虽然新鲜组织的快速细胞尺度成像在临床领域可以具有显着好处，但可以 还赋予基本和翻译的研究用途，用于本质上是组织学 - 病理学或 直接来自实验动物模型的与毒理学相关的图像（在长凳上），并可能有助于营救 这样的研究人员不得不依靠经常劳累过度，有时不可用的传统组织学和 病理服务。