The quantification of nano- and microparticles is critical for diverse applications relying on the exact knowledge of the particle concentration. Although many techniques are available for counting particles, there are some limitations in regards to counting with low-scattering materials and facile counting in harsh organic solvents. Herein, we introduce an easy and rapid particle counting technique, termed "immobilized particle imaging" (IPI), to quantify fluorescent particles with different compositions (i.e., inorganic or organic), structures (i.e., solid, porous, or hollow), and sizes (50-1000 nm) dispersed in either aqueous or organic solutions. IPI is achieved by immobilizing particles of interest in a cell matrix-like scaffold (e.g., agarose) and imaging using standard microscopy techniques. Imaging a defined volume of the immobilized particles allows for the particle concentration to be calculated from the count numbers in a fixed volume. IPI provides a general and facile approach to quantify advanced nano- and microparticles, which may be helpful to researchers to obtain new insights for different applications (e.g., nanomedicine).