Ex Vivo Fusion Confocal Microscopy of Liver Biopsies: Diagnostic Pattern Identification and Correlation with Conventional Microscopy

Ex vivo Fusion Confocal Microscopy (eFuCM) is a promising new technique for real-time histological diagnosis, requiring minimal tissue preparation and avoiding tissue waste. This study aimed to evaluate the feasibility of eFuCM in identifying key liver biopsy lesions and patterns, and to assess the impact of eFuCM reading experience on diagnostic accuracy. Twenty-three fresh liver biopsies were analyzed using eFuCM to produce H&E-like digital images, which were reviewed by two pathologists and compared with a conventional H&E diagnosis. The liver architecture was clearly visible on the eFuCM images. Pathologist 1, with no prior eFuCM experience, achieved a substantial agreement with the H&E diagnosis (κ = 0.65), while Pathologist 2, with eFuCM experience, reached almost perfect agreement (κ = 0.88). However, lower agreement levels were found in the evaluation of inflammation. Importantly, tissue preparation for eFuCM did not compromise subsequent conventional histological processing. These findings suggest that eFuCM has great potential as a time- and material-saving tool in liver pathology, though its diagnostic accuracy improves with pathologist experience, indicating that there is a learning curve related to its use.