The preparation of supramolecular cyclodextrin (CD) assemblies and control of their assembly mode through guest inclusion in CD cavities have been actively studied. Contrarily, there are limited reports on the control of the assembly mode of guest-free CD molecules by external stimuli. Herein, we report the use of 2-O-methylated β-cyclodextrin (2-Me-β-CD) as an effective building block in fabricating supramolecular assemblies with diverse morphologies and molecular arrangements through assembly mode control by various stimuli, such as temperature and solvent. When methanol and diethyl carbonate were used as good and poor solvents, respectively, 2-Me-β-CD formed an amorphous assembly through solvent evaporation on a polyethylene terephthalate (PET) substrate. Increasing the drying temperature and using crystalline substrates, such as highly oriented pyrolytic graphite (HOPG) and sapphire, changed the assembly mode of 2-Me-β-CD to a head-to-tail channel assembly. However, when a 2-Me-β-CD/1-propanol solution was mixed with linear alkanes as a poor solvent, 2-Me-β-CD with head-to-head channel assembly was formed as a precipitate. Additionally, when the corresponding cyclic alkane was used as an alternative poor solvent, an organogel composed of 2-Me-β-CD with head-to-head channel assemblies was obtained. The organogel obtained became a precipitate composed of 2-Me-β-CD with cage-type assembly upon heating at 50 °C. Among the supramolecular assemblies fabricated in this study, the head-to-tail channel assembly is a rare molecular assembly of β-CD and its derivatives. It possesses a modified columnar cavity that has potential applications in molecular recognition and sensing.