The rapid estimation of fat content using spectroscopic-based sensors, irrespective of fish species (salmon, trout, sea bass, sea bream, tuna, cod, and mackerel), was primarily investigated in the present study. The fat content and fatty acid (FA) composition was quantified with reference methods. Fourier-Transform Infrared (FTIR) and Fourier-Transform Near-Infrared (FT-NIR) spectroscopy, along with multispectral imaging (MSI) instruments (both benchtop and portable), were evaluated for their ability to rapidly predict fat content in ground samples using data analysis. The performance of the PLS-R models was evaluated according to root mean square error (RMSE), coefficient of determination (R2) and residual prediction deviation (RPD). Also fish were classified as fat/low-fat and low/medium/high-fat using partial least squares discriminant analysis (PLS-DA). All fish species exhibited a consistent pattern of unsaturated fatty acids (UNFA) > monounsaturated fatty acids (MUFA) > polyunsaturated fatty acids (PUFA) > saturated fatty acids (SFA) and the range of fat content was from 22.8 (salmon) to 0.02 (cod), expressed in g/100 g of sample based on the reference methods. In terms of fat content prediction using rapid sensors the best performance indices of the test set were obtained from the benchtop-MSI (RMSE = 1.475, R2 = 0.847, RPD = 2.581) and FT-NIR (RMSE = 1.638, R2 = 0.855, RPD = 2.651) instruments, while FTIR and portable-MSI had scores of RMSE = 1.874, R2 = 0.815, RPD = 2.309 and RMSE = 1.737, R2 = 0.786, RPD = 2.191, respectively. All sensors discriminated fat from low-fat samples (accuracy = 100%). For low/medium/high fat the best results were achieved by benchtop-MSI (accuracy = 94.12%), followed by portable-MSI (accuracy = 88.24%) and FTIR (accuracy = 84.21%). Results were less satisfactory for the FT-NIR method (accuracy = 68.75%). This study demonstrates that vibrational and multispectral imaging spectroscopies, when coupled with data analysis, have potential to provide information about the nutritional quality (e.g., fat content) independent of the fish species. Such insights could contribute to the automation and potential digitalization of processes within the fields of food nutrition and the seafood industry.