The gelation of a hydrophobically modified hyaluronic acid aqueous solution which shows a lower critical solution temperature of about 25 °C was investigated by multi-particle tracking microrheology. The linear viscoelasticity of the gelling system is converted from the microrheological data. The critical gelling temperature T_gel = 36.3 °C was determined from the loss tangent by the Winter–Chambon criterion. The critical exponent n = 0.62 was determined from the shift factors of the time–cure superposition. The length scales of the dynamic heterogeneity of the gelling system were analyzed using a proposed framework where single-particle and multi-particle non-Gaussian parameters were compared. The length scale of the dynamic heterogeneous regions monotonically decreases during the gelation process, consistent with the nucleation-and-growth mechanism of phase separation. Distributions of local viscosity in the gelling system were extracted from the observed distributions of particle displacement as a time-dependent fingerprint of the dynamic heterogeneity of the gelling system. The results and analyzing methods proposed in the present work can be applied to other microrheological studies.