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

BACKGROUND: Ex vivo confocal laser scanning microscopy (ex vivo CLSM) is a novel diagnostic method allowing rapid, high-resolution imaging of excised skin samples. Furthermore, fluorescent detection is possible using fluorescent-labelled antibodies. OBJECTIVE: To assess the applicability of ex vivo CLSM in the detection of basement membrane (BM) fluorescence in bullous pemphigoid (BP) and to compare its diagnostic accuracy with direct immunofluorescence (DIF) microscopy. METHODS: A total of 81 sections of 49 BP patients with positive DIF microscopy findings were examined using ex vivo CLSM in reflectance and fluorescence mode following staining with fluorescent-labelled IgG and C3 antibodies. RESULTS: Ex vivo CLSM showed an overall performance of 65.3% in identifying BM fluorescence in BP patients. IgG and C3 deposition along the BM was detected in 50% and 45.5% of the patients, respectively. The sensitivity of ex vivo CLSM in detecting BM fluorescence was low (IgG: 50%, C3: 45.5%), but the specificity was high (IgG: 100, C3: 90%). In addition to immunoreactivity, ex vivo CLSM could display subepidermal inflammatory cells similar to histological examination in 84% of patients. CONCLUSIONS: Basement membrane fluorescence could be identified with ex vivo CLSM in the skin sections of BP patients. Ex vivo CLSM enables simultaneous and rapid detection of histopathological and immunofluorescence findings in the same session, albeit with a lower sensitivity than DIF in detecting BM fluorescence.

Evaluation of the response to treatment of psoriasis vulgaris with reflectance confocal microscopy.

BACKGROUND/OBJECTIVE: Reflectance confocal microscopy (RCM) is a noninvasive, objective imaging technique that provides in vivo, high-resolution skin imaging. We sought to assess epidermal and dermal changes associated with the psoriasis and its treatment with RCM before the treatment and at weeks 4 and 8 of the treatment. MATERIAL AND METHODS: This is an investigator-blinded, internal-controlled, follow-up study. A total of 25 patients with plaque psoriasis were included in the study. The RCM evaluation criteria were defined on the basis of the histopathological diagnostic criteria for psoriasis. The clinical severity of the psoriasis was evaluated using the Psoriasis Area Severity Index (PASI). RESULTS: The RCM findings which were correlated with the PASI can be used to follow up the patient's response to treatment have been identified as follows: the acanthosis, the number of spongiotic sites, the number of regular/irregular honeycomb-like sites, the number of epidermal inflammatory cells, the number of focal microabscesses, the total epidermal thickness, the number of nonedge dermal papillae, the length of the papillary dermis, the number of dermal inflammatory cells, and the vascularization in the papillary dermis (P < 0.05). CONCLUSION: This is the first study with a large group of patients to perform a noninvasive assessment with RCM of the response of psoriasis to different treatments: phototherapy, systemic and topical treatment. Micrometric and morphometric changes occurring in the psoriatic tissue during the 8-week treatment period were identified by in vivo RCM in a noninvasive manner. RCM is capable of monitoring of treatment response in psoriasis.