In this paper, we review the application of fluorescent probe technology in self-assembly systems of amphiphile aqueous solutions. Fluorescence technology, especially fluorescence probe technology has been extensively employed to explore local information about various molecular assemblies. The following contents are covered: (1) The critical aggregate concentration, microviscosity, and micropolarity may be obtained fromfluorescence parameters such as emission maxima, fluorescence intensity, and lifetime etc. (2) Probe quenching, especially, time-resolved fluorescence quenching (TRFQ) can give information about aggregation number and surface charge density which can be used as the indication on aggregate transition. (3) Fluorophore-labeled amphiphiles which can take part in the formation of aggregates reveal more precise information, so that an in situ investigation of local environments is possible. Moreover, fluorescence resonance energy transfer (FRET) and excited state proton transfer (ESPT) are also useful in this field. Therefore, fluorescence probe technology provides us with a simple and effective method to study organized amphiphiles systems.