Ex vivo confocal laser scanning microscopy: A diagnostic technique for easy real-time evaluation of benign and malignant skin tumours.

Ex vivo confocal laser scanning microscopy (ex vivo CLSM) is a novel diagnostic tool for a quick bedside evaluation of freshly excised tissue, comparable to histology. We aimed to assess the sensitivity and specificity of ex vivo CLSM in detecting malignant features, to validate its reliability in identifying various skin tumours based on a combination of confocal features and to evaluate the digital staining mode (DS). One-hundred twenty freshly excised skin samples from 91 patients were evaluated. Each lesion was screened for the presence of 23 predefined confocal criteria with ex vivo CLSM, followed by a histopathological examination. The diagnostic agreement between ex vivo CLSM and histology was 89.2%. The diagnostic accuracy of ex vivo CLSM in detecting malignancy reached a sensitivity of 98% and a specificity of 76%. Ex vivo CLSM enabled a rapid identification of the most common skin tumours, the tumour dignity and cytological features. The DS demonstrated a close resemblance to conventional histopathology.

Ex vivo confocal laser scanning microscopy: An innovative method for direct immunofluorescence of cutaneous vasculitis.

Ex vivo confocal laser scanning microscopy (ex vivo CLSM) offers an innovative diagnostic approach through vertical scanning of skin samples with a resolution close to conventional histology. In addition, it enables fluorescence detection in tissues. We aimed to assess the applicability of ex vivo CLSM in the detection of vascular immune complexes in cutaneous vasculitis and to compare its diagnostic accuracy with direct immunofluorescence (DIF) microscopy. Eighty-two sections of 49 vasculitis patients with relevant DIF microscopy findings were examined using ex vivo CLSM following staining with fluorescent-labeled IgG, IgM, IgA, C3 and fibrinogen antibodies. DIF microscopy showed immunoreactivity of vessels with IgG, IgM, IgA, C3 and Fibrinogen in 2.0%, 49.9%, 12.2%, 59.2% and 44.9% of the patients, respectively. Ex vivo CLSM detected positive vessels with the same antibodies in 2.0%, 38.8%, 8.2%, 42.9% and 36.7% of the patients, respectively. The detection rate of positive superficial dermal vessels was significantly higher in DIF microscopy as compared to ex vivo CLSM (P < .05). Whereas, ex vivo CLSM identified positive deep dermal vessels more frequently compared to DIF microscopy. In conclusion, ex vivo CLSM could identify specific binding of the antibodies in vessels and showed a comparable performance to conventional DIF microscopy in diagnosing vasculitis.

Ex vivo confocal microscopy features of cutaneous squamous cell carcinoma.

BACKGROUND: Rapid microscopic evaluation of cutaneous squamous cell carcinoma (SCC), its grade of differentiation and level of invasiveness would enable better management of patients' therapy. OBJECTIVES: Analyzing specific ex vivo confocal microscopy criteria whether they can predict diagnosis of invasive SCC vs carcinoma in situ and poorly differentiated or undifferentiated vs well and moderately differentiated SCC. METHODS: Ex vivo confocal images of 102 SCCs in 57 patients were evaluated immediately after excision for the presence of predefined criteria based on confocal and histological knowledge. RESULTS: In histopathological examination, 30 SCCs were in situ and 72 invasive. Of these, 29 invasive SCC tumors were well, 19 moderately, 15 poorly differentiated and 9 undifferentiated. χ2 analysis demonstrated that presence of erosion/ulceration, plump bright or speckled cells in dermis, keratin pearls and peritumoral inflammatory infiltrate correlated with diagnosis of invasive SCC. Erosion/ulceration and peritumoral inflammatory infiltrate were observed more frequently in poorly differentiated or undifferentiated tumors. Plump bright or speckled cells in the dermis were observed less often in well-differentiated tumors. The presence of keratin pearls was associated with well or moderately differentiated tumors. CONCLUSION: Ex vivo CLSM allowed rapid examination of SCC and provided useful information on invasiveness and grading of the tumor.