Ex vivo fluorescence confocal microscopy for intraoperative, real‐time diagnosis of cutaneous inflammatory diseases: A preliminary study

Abstract

Ex vivo fluorescence confocal microscopy (FCM) is an innovative imaging tool that can be used intraoperatively to obtain real-time images of untreated excised tissue with almost histologic resolution. As inflammatory diseases often share overlapping clinical features, histopathology evaluation is required for dubious cases, delaying definitive diagnoses, and therefore therapy. This study identifies key-features at ex vivo FCM for differential diagnoses of cutaneous inflammatory diseases, in particular, psoriasis, eczema, lichen planus and discoid lupus erythematosus. Retrospective ex vivo FCM and histological evaluations with relevant diagnoses were correlated with prospectively reported histopathologic diagnoses, to evaluate agreement and the level of expertise required for correct diagnoses. We demonstrated that ex vivo FCM enabled the distinction of the main inflammatory features in most cases, providing a substantial concordance to histopathologic diagnoses. Moreover, ex vivo FCM and histological evaluations reached a substantial agreement with histopathologic diagnoses both for all raters and for each operator. After a yet to be defined learning curve, these preliminary results suggest that dermatologists may be able to satisfactorily interpret ex vivo FCM images for correct real-time diagnoses. Despite some limitations mainly related to the equipment of FCM with a single objective lens, our study suggests that ex vivo FCM seems a promising tool in assisting diagnoses of cutaneous inflammatory lesions, with a level of accuracy quite close to that offered by histopathology. This is the first study to investigate ex vivo FCM application in cutaneous inflammatory lesions, and to evaluate the diagnostic capability of this technology.

Ex vivo konfokale Laserscanmikroskopie – die neue Bedside-Histologie

“…Da entzündliche Erkrankungen häufig überlappende klinische Merkmale aufweisen, ist in zweifelhaften Fällen eine histopathologische Bewertung häufig erforderlich, um die endgültigen Diagnosen und damit die Therapie nicht zu verzögern. Eine retrospektive Studie zur Identifizierung der Schlüsselmerkmale für die Differenzialdiagnose von entzündlichen Hauterkrankungen, insbesondere Psoriasis, Ekzem, Lichen planus und diskoidem Lupus erythematodes in der ex vivo KLM analysierte und korrelierte die ex vivo KLM mit histologischen Untersuchungen der relevanten Diagnosen, um die Übereinstimmung und das für die korrekte Diagnose erforderliche Fachwissen zu bewerten [10]. Hier zeigte sich, dass die ex vivo KLM in den meisten Fällen die Unterscheidung der wichtigsten Entzündungsmerkmale ermöglichte und eine erhebliche Übereinstimmung mit den histopathologischen Diagnosen ergab [10].…”

Ex vivo fluorescence confocal microscopy: the first application for real‐time pathological examination of prostatic tissue

“…Preliminary research into FCM as a diagnostic tool for other visceral organ cancers seems promising . Furthermore, it showed to be a promising tool for diagnosis of cutaneous inflammatory skin lesions .…”

Ex vivo confocal laser scanning microscopy for bullous pemphigoid diagnostics: new era in direct immunofluorescence?

“…So far, ex vivo CLSM has been mainly used in the intra‐ and postoperative assessment of skin tumours, the control of surgical skin margins and as an assistive method for Mohs microscopic surgery . A few studies have reported that ex vivo CLSM examination on freshly excised samples may also be helpful in the rapid diagnosis of inflammatory and infectious skin lesions . Recently, our group combined ex vivo CLSM examination with fluorescent‐labelled antibody staining in the tissue sections of patients with melanoma, BCC and cutaneous vasculitis, and showed that ex vivo CLSM may have potential as an alternative method in immunofluorescent examination of skin.…”

“…[2][3][4][5][6][7][8][9][10] A few studies have reported that ex vivo CLSM examination on freshly excised samples may also be helpful in the rapid diagnosis of inflammatory and infectious skin lesions. [11][12][13][14][15] Recently, our group combined ex vivo CLSM examination with fluorescent-labelled antibody staining in the tissue sections of patients with melanoma, BCC 2 and cutaneous vasculitis, 16 and showed that ex vivo CLSM may have potential as an alternative method in immunofluorescent examination of skin.…”