Objective To evaluate the diagnostic performance of probe-based confocal laser endomicroscopy (pCLE) for indeterminate biliary strictures. Methods Twelve patients with indeterminate biliary strictures who underwent pCLE and brush cytology from April 1, 2013 to December 30, 2016 were enrolled. Clinical data, the results of endoscopic retrograde cholangiopancreatography(ERCP), pCLE examination and brush cytology were collected. Compared with post-operative pathology and follow-up over 12 months, sensitivity, specificity, positive predictive value(PPV)，negative predictive value(NPV), accuracy of pCLE and brush cytology of the diagnosis of malignant biliary strictures were analyzed．Results The final diagnosis were 9 malignant and 3 benign. The sensitivity, specificity, PPV, NPV and accuracy of brush cytology were 3/9, 3/3, 3/3 3/9 and 50.0%(6/12), respectively. The corresponding indicators of pCLE were 9/9，2/3，9/10，2/2，and 91.7%(11/12), respectively. Conclusions pCLE shows high sensitivity and accuracy in the diagnosis of malignant biliary strictures，which can be used for differential diagnosis of indeterminate biliary stricture.