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Exploring the utility of Deep Red Anthraquinone 5 for digital staining of ex vivo confocal micrographs of optically sectioned skin
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2020-12-14 , DOI: 10.1002/jbio.202000207
Vinzent Kevin Ortner 1 , Aditi Sahu 2 , Miguel Cordova 2 , Kivanc Kose 2 , Saud Aleissa 2 , Christi Alessi-Fox 3 , Merete Haedersdal 1 , Milind Rajadhyaksha 2 , Anthony Mario Rossi 2
Affiliation  

We investigated the utility of the fluorescent dye Deep Red Anthraquinone 5 (DRAQ5) for digital staining of optically sectioned skin in comparison to acridine orange (AO). Eight fresh‐frozen thawed Mohs discard tissue specimens were stained with AO and DRAQ5, and imaged using an ex vivo confocal microscope at three wavelengths (488 nm and 638 nm for fluorescence, 785 nm for reflectance). Images were overlaid (AO + Reflectance, DRAQ5 + Reflectance), digitally stained, and evaluated by three investigators for perceived image quality (PIQ) and histopathological feature identification. In addition to nuclear staining, AO seemed to stain dermal fibers in a subset of cases in digitally stained images, while DRAQ5 staining was more specific to nuclei. Blinded evaluation showed substantial agreement, favoring DRAQ5 for PIQ (82%, Cl 75%‐90%, Gwet's AC 0.74) and for visualization of histopathological features in (81%, Cl 73%‐89%, Gwet's AC 0.67), supporting its use in digital staining of multimodal confocal micrographs of skin.image

中文翻译:

探索 Deep Red Anthraquinone 5 在光学切片皮肤离体共焦显微照片数字染色中的应用

我们研究了荧光染料深红蒽醌 5 (DRAQ5) 在光学切片皮肤数字染色中的效用,并与吖啶橙 (AO) 进行了比较。将八个新鲜冷冻解冻的莫氏废弃组织标本用 AO 和 DRAQ5 染色,并使用离体共聚焦显微镜在三个波长(488 nm 和 638 nm 荧光,785 nm 反射)下成像。图像被叠加(AO + 反射率、DRAQ5 + 反射率)、数字染色,并由三名研究人员评估感知图像质量 (PIQ) 和组织病理学特征识别。除了核染色之外,AO 似乎还对数字染色图像中一部分病例的真皮纤维进行染色,而 DRAQ5 染色对细胞核更具特异性。盲法评估显示出基本一致,支持 DRAQ5 的 PIQ(82%,Cl 75%‐90%,Gwet 的 AC 0.74)和组织病理学特征的可视化(81%,Cl 73%‐89%,Gwet 的 AC 0.67),支持其用于皮肤多模式共聚焦显微照片的数字染色。图像
更新日期:2020-12-14
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