Herein, in a one-pot procedure, fluorescent silica submicrospheres (FSSMSs) with ordered composite structure were facilely fabricated by the approach of AIE-amphiphile-induced hierarchical order self-assembly. A key factor is the use of an AIE-active amphiphile as both light-emitting source and structure-directing agent. As expected, the fluorescent micelles formed by self-assembly of the AIE-active amphiphile were orderly assembled in the silica networks through noncovalent attachment in the system. The resultant FSSMSs have uniform particle size and morphology, demonstrating excellent properties, such as ordered structure, good monodispersity, excellent photostability, no fluorescence leakage and easy synthesis. The results demonstrate that ordered composite structure in FSSMSs is beneficial for its photostability as well as structural stability. As a test proof, the FSSMSs with a mean size of 0.6 mu m were used to check the integrity of microfiltration membranes with 0.45 mu m pores. A simple and convenient approach of using FSSMSs for assessing the level of membrane integrity was proposed. In addition, the FSSMSs composite materials show unexpectedly excitation-dependent emission properties. The special FSSMSs have promising application in multicolor display and bioimaging.