The photophysics of conjugated polymers has generally teen explained based on the interactions between the component conjugated chrotnophores hi a tangled' chain. However, conjugated chromophores are entities with static and dynamic structural disorder, which directly affects the conjugated polymer photophysics.' t we demonstrate the impact of chain structure torsional disorder on the spectral characteristics for a macrocyclic oligothiophene 1, which is obscured in Conventional linear conjugated chromophores by diverse structural disorders such a those in chtomophore size and shape. We used simultaneous multiple fluorescence parameter measurement for a single molecule and quantum-mechanical calculations. to Show that within the fixed conjugation length across the entire ring an inhoinogeneity from torsional disorder in the structure of 1 plays a crucial role in causing its energetic disorder, which affords the spectral broadening of similar to 220 meV. The torsional disorder in 1 fluctuated on the time scale of hundreds of milliseconds, typically accompanied by spectral drifts on the order of similar to 10 meV. The fluctuations could generate torsionaLdefects and change the electronic structure of 1 associated with the ring symmetry. These findings disclose the fundamental:nature of conjugated chromophore that is the most elementary spectroscopic unit in conjugated polymers and suggest the importance of engineering structural disorder to optimize: polymer-based device photophysics. Additionally, we combined defoctised wide-field fluorescence microscopy and linear dichroism obtained from the simultaneous measurements to show that 1 emits polarized light with a changing polarization direction based on the torsional disorder fluctuations.