The wavelength-dependent steady-state fluorescence data for D4 was additionally recorded (Figure ) to confirm that heterogeneous microenvironments are still present for these molecules. The normalised fluorescence profiles can be seen in Supporting Information Section S8. The REE was still found to be present at an identical chromophore concentration between the samples, with a slight increase in the degree of REE experienced over the untethered trans-PyChal molecules. The difference is attributed to the local increase in PyChal-PyChal interactions for D2-4, where the influence of neighbouring PyChal molecules upon the local microenvironments are much more likely than for the free monomers.