Understanding the factors that have influence on the compactness of calixarene assemblies is important for choosing or designing molecules for specific function. In this paper, we comparatively investigated the packing performances of a series of amphiphilic calixarene assemblies by fluorescence anisotropy. Calixarene frameworks, the upper-rim decoration of head groups and the length of lower-rim alkyl chains of amphiphilic calixarenes all affect their packing compactness. This work has guiding significance for further application studies.