Before performing immobilization on the established microcarriers' surface, with TiFe/Au or TiFe/SiO2 coatings, we use in situ Atomic Force Microscopy (in situ AFM) in 1 mM PBS at pH 7.4 in 2D and 3D to confirm the conformation and immobilization efficacy of three varieties of cargo-loaded Psomes on different surfaces. The schematic of the optimized surface immobilization is shown with its respective in situ AFM results demonstrated in Figure . The specific steps of the immobilization procedure on two distinct substrates, namely silicon dioxide (SiO2) and gold (Au) layer, along with their corresponding in situ AFM images, are found in Section 5.2 of the SI (schematic of the immobilization process in Figure ; in situ AFM measurements for SiO2 immobilized surface in Figure ). The physical interaction between the surface and the loaded-Psomes is the same for both substrates, via physical interactions between the βCD cavities (chemically and physically conjugated to the microcarrier surface) and the PEG chains (Psomes surface). We choose gold as it not only exhibits a uniform and high-quality surface covering but also serves as a contrast agent for photoacoustic imaging. This property can potentially enable the tracking of microcarriers in in vivo applications in the future. Moreover, SiO2 is selected as a surface coating because of its biocompatibility and ability to provide the necessary functional group for binding carboxylic groups, which is crucial for the subsequent binding of requested amino-terminated βCD. Both non-covalent conjugation strategies of loaded-Psomes outlined following results: SiO2 layer does not exhibit uniform coverage of cargo-loaded Psomes. When cargo-loaded Psomes are immobilized on a SiO2 surface as opposed to an Au surface, the Psomes are more likely to aggregate and form a non-uniform coating, according to the in-situ AFM images (Figure and Figure ; considering the height amplitudes for scale bar with 400 nm). Subsequently, the coating with prior physisorption with thiol-terminated βCD on the Au demonstrates a superior and more homogeneous Psomes coating that can be chosen for the immobilization of cargo-loaded Psomes on microcarriers with TiFe/Au coating of the outer surface (Figure ), thereby facilitating future photoacoustic tracking.