This review provided a detailed overview of the different synthesis and characterization methods of polymeric nanoparticles. Nanoparticles are defined as solid and colloidal particles of macromolecular substances ranging in size under 100 nm. Different types of nanoparticles are used in many biological fields (bio-sensing, biological separation, molecular imaging, anticancer therapy, etc.). The new features and functions provided by nano dimensions are largely different from their bulk forms. High volume/surface ratio, improved resolution and multifunctional capability make these materials gain many new features.