The physicochemical properties of CSNPs are crucial for understanding their stability, structural integrity, and potential applications in biomedicine. The present chapter examines analytical approaches used to quantify CSNPs, focusing on particle dimensions and size distribution through dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), and other relevant microscopic techniques. The AFM enables imaging at atomic resolution, facilitating the evaluation of surface roughness and nanostructures. SEM and TEM are employed for morphological characterization of particles, focusing on particle shape, uniformity, and structure. The discussion also includes zeta potential results to assess surface charge and colloidal stability in physiological medium. The characterization techniques described establish a systematic protocol linking NP characteristics to their functional performance, thereby enhancing reproducibility and enabling the rational design of CS-based nanocarriers.