The milk protein casein (Cas) has been employed as carbon resource material to synthesize nitrogen-doped carbon dots (N-CDs) via microwave exposure. The dots, when exposed to UV light, produced blue fluorescence. The N-CDs were characterized by ultra violet (UV) spectroscopy, Fourier transformation infrared spectroscopy, X-ray diffraction (XRD), dynamic light scattering analysis, fluorescent microscopy (FM), and transmission electron microscopy (TEM). The XRD analysis revealed a broad peak at 2 theta = 20 degrees, thus indicating the turbostratic carbon phase. TEM analysis and particle size distribution curve revealed that nearly, 85% of the particles had diameter below 10 nm and the particles had spherical geometry. The HRTEM analysis revealed that carbon dots exhibited lattice fringes with a d-spacing of 0.21 nm, corresponding to the (100) plane lattice of graphite. The fluorescence spectral studies indicated a red shift in the emission peak from 420 to 450 nm as the excitation wavelength increased from 300 to 340 nm. The zeta potential of particles was found to be -11.3 mV. Finally, impregnation of N-CDs was studied in Spinacia oleracea leaf. It was observed that as the concentration of N-CDs' solution increased, percent insertion (PI) also increased, but the time required for maximal insertion decreased with increasing concentrations of N-CDs in the feed solutions. In the carbon dots' solution with a concentration of 200 ppm, maximum percent insertion (MPI) was obtained after 80 min. However, with the increasing concentration of N-CDs in the feed solutions, time of getting MPI reduced, i.e., in 600 ppm, it was 30 min, and in 800 ppm, it was 10 min.