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

病理学-确定疾病的原因和影响，通常通过使用 显微镜检查病人的组织-是必要的，以达到正确的癌症诊断，但它有 提交通常微小的组织样本的相关部分用于DNA和其他检测变得越来越重要。 分子和功能测试。确保提交的材料确实含有肿瘤， 数量，并不总是容易的，有时只是准备传统的显微镜载玻片可以消耗最 甚至是整个样本我们开发了一种新的，简单而廉价的方法，我们称之为MUSE， 带UV表面激发的显微镜，可以直接快速地从新鲜样品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标本，用于下游的生物库 分子和功能分析。 MUSE通过确保获得的小活检标本确实适合目的，可以避免 需要让患者在稍后的日期返回进行额外的活检。此外，该方法将允许病变 在更大的组织横截面上的检测，例如器官整体安装， 常规仪器。最后，我们期望MUSE将产生额外的生物信息成像 通过避免传统组织处理和切片中固有的伪影。我们在此着重于促进 基于组织的分子研究、用于异种移植物和培养球状体的活肿瘤收获物以及生物库， 并包括关键的验证研究，以确保MUSE过程不会损害样品效用， 暴露于嵌入染料和/或UV光可能产生的影响。 MUSE依赖于两种机制：1）由于微米-纳米粒子的存在，荧光染料的表面限制激发。 亚300纳米紫外光的尺度渗透;和2）许多常规染料在这种情况下激发的事实 方式发射可见光。这些信号足够明亮，可以被传统的彩色摄像机检测到， 次秒曝光时间，允许大面积快速成像。 MUSE无需福尔马林固定、石蜡 包埋或薄切片。它不需要激光、共焦、多光子或光学相干断层扫描 仪器，最终成本可能在几千美元的范围内，因此在市场上是负担得起的。 点护理（活检点）。使用熟悉的组织学染色剂在几秒钟内对MUSE样品进行染色， 作为曙红和DAPI，以及由此产生的高分辨率图像从荧光转换为H& E样 用于真实的时间解释的明场外观（但具有新颖的和潜在有用的特征） 由病理学家解释。 虽然新鲜组织的快速细胞级成像在临床领域具有显著的益处，但它可以 还授权基本和转化研究用于基本上即时组织学，病理学或 毒理学相关的图像直接从实验动物模型-在工作台上-并可能有助于缓解 这样的研究人员不得不依靠经常过度劳累，有时无法获得，传统的组织学， 病理学服务。

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.