Allometric scaling of body size versus growth rate and mortality has been suggested to be a universal macroecological pattern, as described by the metabolic theory of ecology (MTE). However, whether such scaling generally holds in natural assemblages remains debated. Here, we test the hypothesis that the size-specific growth rate and grazing mortality scale with the body size with an exponent of −1/4 after temperature correction, as MTE predicts. To do so, we couple a dilution experiment with the FlowCAM imaging system to obtain size-specific growth rates and grazing mortality of natural microphytoplankton assemblages in the East China Sea. This novel approach allows us to achieve highly resolved size-specific measurements that would be very difficult to obtain in traditional size-fractionated measurements using filters. Our results do not support the MTE prediction. On average, the size-specific growth rates and grazing mortality scale almost isometrically with body size (with scaling exponent ∼0.1). However, this finding contains high uncertainty, as the size-scaling exponent varies substantially among assemblages. The fact that size-scaling exponent varies among assemblages prompts us to further investigate how the variation of size-specific growth rate and grazing mortality can interact to determine the microphytoplankton size structure, described by normalized biomass size spectrum (NBSS), among assemblages. We test whether the variation of microphytoplankton NBSS slopes is determined by (1) differential grazing mortality of small versus large individuals, (2) differential growth rate of small versus large individuals, or (3) combinations of these scenarios. Our results indicate that the ratio of the grazing mortality of the large size category to that of the small size category best explains the variation of NBSS slopes across environments, suggesting that higher grazing mortality of large microphytoplankton may release the small phytoplankton from grazing, which in turn leads to a steeper NBSS slope. This study contributes to understanding the relative importance of bottom-up versus top-down control in shaping microphytoplankton size structure