These approaches enhance the obtained data by integrating AFM (topography, mechanics) with information from other techniques within the same region of interest. It provides key insights into the structural and biomechanical behaviour of biological systems in their native state and physiological environments, enabling a deeper understanding of complex processes such as biomolecule imaging and cell mechanics. AFM’s versatility is further enhanced through its combination with other techniques, such as Raman spectroscopy, fluorescence microscopy, and scanning electron microscopy (SEM). While Raman spectroscopy provides precise chemical characterization, fluorescence microscopy enables the specific localization of desired interactions. Moreover, the correlative AFM-in-SEM enables precise in-situ data correlation, combining nanoscale topography with different information from SEM channels. Additionally, advanced AFM techniques, such as Fluid AFM, enable nanomanipulation, including single-cell nano-injection, expanding its applications in biomedical research. These combined methodologies provide a deeper understanding across a wide range of life sciences, including cellular processes, biomaterial interactions, and disease-related mechanical alterations, making AFM an excellent and useful tool for modern biological and biomedical research.