Single-molecule defocused wide-field imaging (DWFI) has been demonstrated to be useful to determine molecular structure parameters, such as the orientations of transition dipole moments and the angular relationships between chromophores in multichromophoric molecular systems. For a series of acetylene-linked perylene bisimide (PBI) macrocycles with different ring size comprising three to six PBI dyes, we reconstructed the molecular structure of the multichromophoric system using DWFI method. Furthermore, we revealed that the structural heterogeneities and distortions depend on the ring size. Our findings illustrate the use of DWFI to gain deeper insight into the structure-property relationships of artificial light-harvesting molecular systems.
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