Abstract Fluorescence imaging is a widely used optical technique in biomedical sciences to examine biological specimens with high‐resolution and cellular‐level molecular specificity, including cancer biopsy. However, light scattering caused by heterogeneous structures within the sample makes it difficult to image deeper tissue layers. Refractive index mismatches between tissue components are the primary cause of scattering, limiting the imaging depth. To address this issue and enhance light penetration into the sample, tissue optical clearing techniques were developed, which reduce tissue heterogeneity, thereby minimizing light scattering and improving the depth of light penetration. Several advancements in optical clearing procedures have been realized in recent years for applications in biological research. Though there exist several reviews on optical clearing techniques, none of them have focused on their application in fluorescence imaging. This review examines various studies that have combined optical clearing methods with fluorescence imaging, with a focus on the imaging techniques employed and its diagnostic and pathological applications. The origins of fluorescence signals and preservation strategies are also discussed in detail.
Pramod et al. (Mon,) studied this question.