The dynamics and fate of endosomal vesicles following endocytosis at the plasma membrane is central to understanding of transport phenomena and signal processing by living cells. Following internalization, cargo localizes in newly generated vesicles at the plasma membrane and subsequently within Rab5 positive endosomal compartments. Transient perturbations that drive rapid redistribution of proteins or short-lived interactions can only be observed by rapid imaging. Using lattice light sheet microscopy based rapid volumetric imaging; We elucidate the complex relations between the dynamics of endosomes marked by APPL1, EEA1 and Rab5 in non-cargo steady state conditions as well as the first time points of transferrin and EGFR entry into vesicles. We find that while transferrin does not alter the ‘state’ of endosomal dynamics, EGFR actuates a subset of endosomes and alters the endosomal trafficking characteristics.