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The present work describes the preparation of nano-lignin particles from calcium lignosulfonate (CL). The nano-lignin was fabricated from colloidal lignin–polyacrylamide complexes via self-assembly. The sizes of the nano-lignin particles were examined by dynamic light scattering (DLS) and scanning electron microscopy (SEM). The results indicated that the average particle size of the prepared nano-lignin was approximately 100 nm. In addition, the obtained nano-lignin exhibited enhanced fluorescence intensity when compared with the original lignin, which might represent a potential application of this nano-particle product.