Acoustic coupling agents serve as critical interfacial materials connecting piezoelectric transducers with microfluidic chips in acoustofluidic systems. Their performance directly impacts acoustic wave transmission efficiency, device reusability, and reliability in biomedical applications. Considering the rapidly growing body of research in the field of acoustic microfluidics, this review aims to serve as an all-in-one reference on the role of acoustic coupling agents and relevant considerations pertinent to acoustofluidic devices for anyone working in or seeking to enter the field of disposable acoustofluidic devices. To this end, this review seeks to summarize and categorize key aspects of acoustic couplants in the implementation of acoustofluidic devices by examining their underlying physical mechanisms, material classifications, and core applications of coupling agents in acoustofluidics. Gel-based coupling agents are particularly favored for their long-term stability, high coupling efficiency, and ease of preparation, making them integral to acoustic flow control applications. In practice, coupling agents facilitate microparticle trapping, droplet manipulation, and biosample sorting through acoustic impedance matching and wave mode conversion (e.g., Rayleigh-to-Lamb waves). Their thickness and acoustic properties (sound velocity, attenuation coefficient) further modulate sound field distribution to optimize acoustic radiation forces and thermal effects. However, challenges remain regarding stability (evaporation, thermal degradation) and chip compatibility. Further aspects of research into gel-based agents requiring attention include multilayer coupled designs, dynamic thickness control, and enhancing biocompatibility to advance acoustofluidic technologies in point-of-care diagnostics and high-throughput analysis.