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首页> 外文OA文献 >DRAQ5 and Eosin (‘DE’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues
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DRAQ5 and Eosin (‘DE’) as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues

机译：DRAQ5和曙红（'D＆E'）作为苏木精和曙红的类似物，用于新鲜组织的快速荧光组织学

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Real-time on-site histopathology review of biopsy tissues at the point-of-procedure has great potential for significant clinical value and improved patient care. For instance, on-site review can aid in rapid screening of diagnostic biopsies to reduce false-negative results, or in quantitative assessment of biospecimen quality to increase the efficacy of downstream laboratory and histopathology analysis. However, the only currently available rapid pathology method, frozen section analysis (FSA), is too time-and labor-intensive for use in screening large quantities of biopsy tissues and is too destructive for maximum tissue conservation in multiple small needle core biopsies. In this work we demonstrate the spectrally-compatible combination of the nuclear stain DRAQ5 and the anionic counterstain eosin as a dual-component fluorescent staining analog to hematoxylin and eosin intended for use on fresh, unsectioned tissues. Combined with optical sectioning fluorescence microscopy and pseudo-coloring algorithms, DRAQ5 and eosin ("D&E") enables very fast, non-destructive psuedohistological imaging of tissues at the point-of-acquisition with minimal tissue handling and processing. D&E was validated against H&E on a one-to-one basis on formalin-fixed paraffin-embedded and frozen section tissues of various human organs using standard epi-fluorescence microscopy, demonstrating high fidelity of the staining mechanism as an H&E analog. The method was then applied to fresh, whole 18G renal needle core biopsies and large needle core prostate biospecimen biopsies using fluorescence structured illumination optical sectioning microscopy. We demonstrate the ability to obtain high-resolution histology-like images of unsectioned, fresh tissues similar to subsequent H&E staining of the tissue. The application of D&E does not interfere with subsequent standard-of-care H&E staining and imaging, preserving the integrity of the tissue for thorough downstream analysis. These results indicate that this dual-stain pseudocoloring method could provide a real-time histology-like image at the time of acquisition and valuable objective tissue analysis for the clinician at the time of service.
机译：在过程点对活检组织进行实时现场组织病理学检查具有巨大的潜在临床价值和改进的患者护理。例如，现场审查可以帮助快速筛查诊断性活检以减少假阴性结果，或者有助于定量评估生物样本质量以提高下游实验室和组织病理学分析的效率。然而，当前唯一可用的快速病理学方法，冷冻切片分析（FSA），对于筛查大量活检组织太费时费力，并且对于多次小针芯活检中最大程度的组织保存来说也具有破坏性。在这项工作中，我们展示了核染色剂DRAQ5和阴离子复染曙红的光谱兼容组合物，作为苏木精和曙红的双组分荧光染色类似物，旨在用于新鲜的，未切开的组织。结合光学切片荧光显微镜和伪着色算法，DRAQ5和曙红（“ D＆E”）可以在采集点对组织进行非常快速的非破坏性伪组织成像，而对组织的处理和处理最少。使用标准的落射荧光显微镜在各种人体器官的福尔马林固定石蜡包埋的和冷冻的切片组织上一对一地对D＆E进行了H＆E验证，证明了H＆E类似物染色机制的高保真度。然后使用荧光结构照明光学切片显微镜将该方法应用于新鲜的，完整的18G肾针核心活检和大针核心前列腺活检。我们证明了获得未切开的，新鲜组织的高分辨率组织学样图像的能力，类似于组织的后续H＆E染色。 D＆E的应用不会干扰随后的护理标准H＆E染色和成像，从而保留了组织的完整性以进行彻底的下游分析。这些结果表明，这种双色伪彩色方法可以在采集时提供实时的组织学样图像，并为临床医生在服务时提供有价值的客观组织分析。

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