Photosynthetic microalgae not only perform fixation of carbon dioxide but also produce valuable byproducts such as lipids and pigments. However, due to the lack of effective tools for rapid and noninvasive analysis of microalgal cellular contents, the efficiency of strain screening and culture optimizing is usually quite low. This study applied single-cell electrorotation on Scenedesmus abundans to assess cellular dielectric properties during lipid accumulation and to promptly quantify total cellular contents. The experimental electrorotation spectra were fitted with the double-shell ellipsoidal model, which considered varying cell wall thickness, to obtain the dielectric properties of cellular compartments. When the amount of total lipids increased from 15.3 wt% to 33.8 wt%, the conductivity and relative permittivity of the inner core (composed of the cytoplasm, lipid droplets, and nucleus) decreased by 21.7% and 22.5%, respectively. These dielectric properties were further used to estimate the total cellular lipid contents by the general mixing formula, and the estimated values agreed with those obtained by weighing dry biomass and extracted lipids with an error as low as 0.22 wt%. Additionally, the conductivity and relative permittivity of cell wall increased during nitrogen-starvation conditions, indicating the thickenin