Imaging and informatics techniques to spatially map tumor-associated collagen: novel cancer diagnostic tools (2 of 2)

项目摘要：病理学-确定疾病的原因和后果，通常通过使用 显微镜检查患者的组织--对于得出正确的癌症诊断是必不可少的，但它 提交通常很小的组织样本的相关部分用于DNA和其他 分子和功能测试。确保提交的材料中确实含有足够的肿瘤 数量，并不总是容易的，有时仅仅是准备常规的显微镜载玻片就可以消耗最多的 甚至是整个标本。我们开发了一种新的、简单且廉价的方法，我们称之为缪斯，用于 带有UV表面激发的显微镜，可以直接快速地从新鲜的图像中提供高分辨率图像 组织而不消耗它，因此可以保存完整的、高质量的标本，用于下游的生物库 分子和功能分析。 缪斯，通过确保获得的小样本确实适合目的，可以避免 需要让病人在晚些时候回来做额外的活组织检查。此外，该方法将允许损伤 检测更大的组织横截面，如器官整体支架，目前不适用于 常规仪器。最后，我们预计缪斯将产生额外的生物信息成像。 通过避免常规组织处理和切片中固有的伪影。我们在这里关注的是促进 基于组织的分子研究，异种移植和培养球体的活肿瘤收获，以及生物库， 并包括关键的验证研究，以确保缪斯过程不会损害样本效用 暴露在插层染料和/或紫外线下可能产生的影响。 MUSE依赖于两种机制：1)微米级荧光染料的表面受限激发。 亚300纳米紫外光的比例穿透；以及2)许多传统染料在这方面的激发 发出可见光的方式。这些信号足够亮，可以由传统的彩色摄像机使用 亚秒级曝光时间，可实现大面积快速成像。 MUSE消除了对福尔马林固定、石蜡切片等常规组织学处理的任何要求 嵌入法，或薄切法。它不需要激光、共焦、多光子或光学相干层析成像 仪器，最终成本可能在几千美元的范围内，因此负担得起 护理点(活检点)。缪斯的样本在几秒钟内就可以用熟悉的组织学染色进行染色，比如 作为曙红和DAPI，并由此产生的高分辨率图像从荧光转换为类H&E Brightfield外观，便于实时口译(但具有新颖且潜在有用的功能) 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有显著的好处，但它可以 还支持基础和转化性研究用于基本上即时的组织学、病理学或 毒理学--直接来自实验动物模型的相关图像--在工作台上--可能有助于缓解 这样的研究人员不得不依赖经常超负荷工作，有时无法获得的传统组织学和 病理服务。

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.