We use Fourier-Bessel Image Decomposition (FBID) of microscopy images to investigate the size, refractive index and 3-dimensional position of individual colloidal microspheres. With measurements of monodisperse polystyrene and poly(methyl methacrylate) particles we achieve a resolution of 1% in size and 0.2% in refractive index for a single image which is sufficient for accurate in situ characterization of polydisperse colloids. Also the binding of avidin molecules to individual biotinylated polystyrene particles is resolved. Finally, the FBID method offers a straightforward approach to 3-dimensional out-of-focus tracking. Here, the z-position of a freely diffusing particle is calculated by applying the statistics of Brownian motion to its set of Fourier-Bessel coefficients.