The thermal aggregation of lysozyme has been analyzed in water/ethanol solutions at low pH to induce the specific protein aggregation pathway which leads to fibrillar structures in a few hours. In this solvating medium, the protein undergoes a conformational rearrangement promoting the formation of fibrils that are structurally similar to amyloid ones. As the process evolves with different steps, a multitechnique approach has been used by means of analytical probes that can be selectively sensitive in the detection of the different stages of protein association. Fourier transform infrared spectroscopy, intrinsic fluorescence, stationary fluorescence anisotropy, transmission electron microscopy (TEM), and atomic force microscopy (AFM) measurements have been carried out at different times to access and characterize the whole aggregation pathway. The data recorded with different experimental setups revealed different sensitivity to different stages of protein assembling. The whole set of data together with the direct visualization of different aggregate structures by use of TEM and AFM imaging enable to discuss a possible mechanism of fibrillation.