It is recognized that water migration and redistribution, starch retrogradation and gluten transformation are one of the main causes of bread staling which can be defined as loss of freshness of bread. Since water migration and redistribution during bread staling are considered as the main factors to affect bread staling, a number of studies on water dynamics in bread staling have been reported. However, staling mechanism is a complex subject, which cannot be fully and easily understood. There are many methods to investigate and analyze the bread staling mechanism. It is very important to analyze the bread staling phenomenon with appropriate analytical tools and, moreover, combination and interpretation of multiple instrumental analyses are more important to understand bread staling mechanism. Therefore, in this review, analytical methodologies using certain instruments, such as Differential Scanning Calorimetry, Thermomechanical Analysis and Nuclear Magnetic Resonance, for bread staling are summarized and discussed.