DRAQ5 and Eosin ('D&E') as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues.
DRAQ5 and Eosin ('D&E') as an Analog to Hematoxylin and Eosin for Rapid Fluorescence Histology of Fresh Tissues.
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DOI：
10.1371/journal.pone.0165530
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Brown JQ
中科院分区：
文献类型：
--
作者：
Elfer KN;Sholl AB;Wang M;Tulman DB;Mandava SH;Lee BR;Brown JQ
在手术点对活检组织进行实时现场组织病理学检查具有巨大的临床价值和改善患者护理的潜力。例如，现场审查可以帮助快速筛查诊断活检，以减少假阴性结果，或定量评估生物标本质量，以提高下游实验室和组织病理学分析的效率。然而，目前唯一可用的快速病理学方法，冷冻切片分析（FSA），对于用于筛选大量活检组织来说太费时费力，并且对于多个小针芯活检中的最大组织保存来说太破坏性。在这项工作中，我们证明了光谱兼容的组合的核染色剂DRAQ5和阴离子复染曙红作为双组分荧光染色类似物的苏木精和曙红用于新鲜的，未切片的组织。结合光学切片荧光显微镜和伪着色算法，DRAQ5和曙红（“D&E”）能够在采集点以最小的组织处理和加工实现组织的非常快速、非破坏性的伪组织学成像。使用标准落射荧光显微术，在福尔马林固定的石蜡包埋和冷冻切片组织的各种人体器官上，一对一地针对H&E验证D&E，证明了作为H&E类似物的染色机制的高保真度。然后，该方法应用于新鲜的，整个18 G肾针芯活检和大针芯前列腺生物标本活检，使用荧光结构照明光学切片显微镜。我们证明了获得类似于随后的组织H&E染色的未切片的新鲜组织的高分辨率组织学样图像的能力。D&E的应用不会干扰后续的标准护理H&E染色和成像，从而保持组织的完整性以进行彻底的下游分析。这些结果表明，这种双染色假彩色方法可以在采集时为临床医生提供实时组织学样图像，并在服务时为临床医生提供有价值的客观组织分析。
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.
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影响因子：
3.7
作者：
Giacomelli MG;Husvogt L;Vardeh H;Faulkner-Jones BE;Hornegger J;Connolly JL;Fujimoto JG
通讯作者：
Fujimoto JG
影响因子：
3.7
作者：
Fu HL;Mueller JL;Whitley MJ;Cardona DM;Willett RM;Kirsch DG;Brown JQ;Ramanujam N
通讯作者：
Ramanujam N
影响因子：
3.6
作者：
Leigh SY;Liu JT
通讯作者：
Liu JT
影响因子：
3.8
作者：
Dobbs, Jessica;Krishnamurthy, Savitri;Richards-Kortum, Rebecca
通讯作者：
Richards-Kortum, Rebecca
影响因子：
2.2
作者：
Gareau, Daniel S.;Jeon, Hana;Rajadhyaksha, Milind
通讯作者：
Rajadhyaksha, Milind