Evaluating Ex Vivo Fluorescence Confocal Microscopy for Intraoperative Diagnosis in Pediatric Surgery: A Feasibility Study

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Ex vivo fluorescence confocal microscopy (FCM) is an emerging technology that enables real-time, high-resolution digital imaging of freshly excised tissues without requiring standard histological preparation. This study aims to evaluate the diagnostic performance of FCM compared with conventional histology in a pediatric population with suspected oncological pathology.A total of 18 tissue samples from pediatric patients with suspected oncological lesions were analyzed using FCM. The results were compared with the definitive diagnoses obtained via conventional histology, serving as the gold standard. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated to assess the diagnostic accuracy. Cohen's kappa coefficient (Κ), 95% confidence interval (95% CI), and p-value were computed to evaluate the interobserver agreement and the concordance between FCM and histology.FCM correctly identified 10 positive cases (true positives) and 7 negative cases (true negatives), with 1 false positive and no false negatives. Interobserver agreement was perfect, with a Cohen's kappa coefficient of 1.00 (95% CI: 0.81–1.00, p

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Article · Aug 2025 · 1 min read

In brief

This feasibility study demonstrates that ex vivo fluorescence confocal microscopy can provide rapid, accurate intraoperative tissue diagnosis in pediatric oncology cases without traditional histological processing. Analyzing 18 samples, FCM showed high diagnostic accuracy with perfect interobserver agreement, correctly identifying malignant and benign tissues in real-time during surgery.

Ex vivo FCM achieved 100% sensitivity and 87.5% specificity for pediatric oncological diagnosis with perfect interobserver agreement (κ=1.00).
FCM correctly identified all malignant cases (10/10) with no false negatives, critical for intraoperative surgical margin assessment.
Technology enables real-time tissue diagnosis without standard histological preparation, potentially reducing intraoperative wait times.
Single false positive (1/18) suggests FCM may overcall benign lesions; confirmatory frozen section may still be needed in select cases.
Feasibility demonstrated in pediatric population; FCM shows promise as adjunct tool for immediate intraoperative pathology guidance.

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