Department of Dermatology, Stanford University School of Medicine, Stanford, CA, USA.
Department of Dermatology and Allergy, University Hospital, LMU Munich, Germany.
Exp Dermatol. 2021 May;30(5):684-690. doi: 10.1111/exd.14265. Epub 2021 Jan 21.
Ex vivo confocal laser scanning microscopy (CLSM) offers real-time examination of excised tissue in reflectance, fluorescence and digital haematoxylin-eosin (H&E)-like staining modes enabling application of fluorescent-labelled antibodies. We aimed to assess the diagnostic performance of ex vivo CLSM in identifying histopathological features and lupus band test in cutaneous lupus erythematosus (CLE) with comparison to conventional histopathology and direct immunofluorescence (DIF). A total of 72 sections of 18 CLE patients were stained with acridine orange (AO), anti-IgG, anti-IgM and anti-IgA; 21 control samples were stained with AO. Subsequently, ex vivo CLSM examination of all samples was performed in reflectance, fluorescence and digital H&E-like staining modes. Superficial and deep perivascular inflammatory infiltration (94.4%), interface dermatitis (88.9%), spongiosis (83.3%) and vacuolar degeneration (77.7%) were the most common features detected with ex vivo CLSM. Kappa test revealed a level of agreement ranging within "perfect" to "good" between ex vivo CLSM and conventional histopathology. ROC analysis showed that the combination of perivascular infiltration, interface dermatitis and spongiosis detected by ex vivo CLSM has the potential to distinguish between CLE and controls. Basement membrane immunoreactivity with IgG, IgM and IgA was identified in 88.8% (n = 15), 55.5% (n = 10) and 55.5% (n = 10) of the CLE samples using ex vivo CLSM, respectively, whereas DIF showed IgG, IgM and IgA positivity in 94.4% (n = 17), 100% (n = 18) and 88.9% (n = 16) of patients, respectively. In conclusion, ex vivo CLSM enables simultaneous histopathological and immunofluorescence examination in CLE showing a high agreement with conventional histopathology, albeit with a lower performance than conventional DIF.
离体共聚焦激光扫描显微镜（CLSM）可实时检查反射、荧光和数字苏木精-伊红（H&E）样染色模式下的离体组织，从而应用荧光标记的抗体。我们旨在评估离体 CLSM 在识别皮肤红斑狼疮（CLE）的组织病理学特征和狼疮带试验方面的诊断性能，并与常规组织病理学和直接免疫荧光（DIF）进行比较。共对 18 例 CLE 患者的 72 个切片进行吖啶橙（AO）、抗 IgG、抗 IgM 和抗 IgA 染色；21 个对照样本进行 AO 染色。随后，在反射、荧光和数字 H&E 样染色模式下对所有样本进行离体 CLSM 检查。最常见的特征是浅表和深部血管周围炎性浸润（94.4%）、界面性皮炎（88.9%）、海绵形成（83.3%）和空泡变性（77.7%）。Kappa 检验显示离体 CLSM 与常规组织病理学之间的一致性程度在“极好”到“良好”之间。ROC 分析表明，离体 CLSM 检测到的血管周围浸润、界面性皮炎和海绵形成的组合具有区分 CLE 和对照组的潜力。在使用离体 CLSM 时，分别在 88.8%（n=15）、55.5%（n=10）和 55.5%（n=10）的 CLE 样本中检测到 IgG、IgM 和 IgA 的基底膜免疫反应，而 DIF 显示 94.4%（n=17）、100%（n=18）和 88.9%（n=16）的患者 IgG、IgM 和 IgA 阳性。总之，离体 CLSM 可同时进行 CLE 的组织病理学和免疫荧光检查，与常规组织病理学具有高度一致性，尽管性能略低于常规 DIF。
