A line of transect was drawn from 1 μm outside of the cell to the nucleolus. The location of the transect line was varied to ensure equidistance from the extracellular space to the nucleolus in all the time courses. Spectral emissions were isolated by selecting regions along the line of transect at 1 μm increments. Graphs were created by utilising small cursors (size = 0.005) to select λmax and widths on the phasor plot corresponding to the cellular region. (A) Depicts smaller λmax values in the extracellular space and near the cytoplasmic region compared to intracellular regions. (B) Depicts a decrease in the width of emissions with respect to a decrease in the distance from the nucleolus. EC = 1 μm outside of cell, CM = cytoplasmic membrane, and NL = nucleolus.</p
