Standard analytical methods for fish freshness assessment are based on the measurement of chemical and physical attributes related to fish appearance, color, meat elasticity or texture, odor, and taste. These methods have plenty of disadvantages, such as being destructive, expensive, and time consuming. All these techniques require highly skilled operators. In the last decade, rapid advances in the development of novel techniques for evaluating food quality attributes have led to the development of non-invasive and non-destructive instrumental techniques, such as biosensors, e-sensors, and spectroscopic methods. The available scientific reports demonstrate that all these new techniques provide a great deal of information with only one test, making them suitable for on-line and/or at-line process control. Moreover, these techniques often require little or no sample preparation and allow sample destruction to be avoided.