A major waste byproduct of oil sands in situ extraction is oil sands tailings, which are a mixture of water, clay, and residual bitumen. These tailings represent a huge ecological footprint in the form of tailings ponds, which not only render large land areas unusable but also prevent reuse of water. The slow dewatering of the tailings ponds poses a major challenge to the industry. The presence of complex inorganic-organic bitumen-clay mixtures in these tailings contributes to this problem. Hence, understanding the nature of the bitumen-clay association and the effect of bitumen on clay particle particle interactions is important for the development of more effective chemicals or processes to accelerate particle aggregation and sedimentation during dewatering. Previous studies that investigate these interactions used techniques that are sensitive only toward inorganic clay but not sensitive towards organic bitumen. Here, we use a high-resolution total internal reflection fluorescence (TIRF) microscopy to help identify the accurate location and distribution of bitumen in mature fine tailings (MFT) samples. We report the first adaptation of TIRF beyond cell biology for visualization of bitumen and its interaction with clay. The high signal-to-noise ratio of TIRF microscopy and a high contrast between the clay and residual bitumen provide images that reveal a wealth of information about the bitumen coverage on clay as well as clay-clay aggregates and how the bitumen positions itself within these aggregates. These images confirm the presence of hydrophobic fine clay agglomerates along with the hydrophilic clay particles in MFT. It is also observed that bitumen coats clay particles, bridges clay agglomerates, and is mostly absent as free bitumen in the bulk of the MFT sample. Our work paves the way for the use of nanophotonic tools in oil sands imaging and provides strategic suggestions for the development of better methods for clay sedimentation and bitumen recovery.