Due to the widespread existence of cell heterogeneity and diversity, the analysis of trace elements and their species in single cell is of great significance to metallomics study. Inductively coupled plasma mass spectrometry (ICP-MS) is a powerful method for trace element quantification. With the data acquisition mode of time-resolved analysis (TRA), ICP-MS can be further used for cells counting and elemental/biomolecular quantification in single cells. The challenge of this strategy exists in sensitivity, selectivity, throughput, and ability in distinguishing individual cells. Various microfluidic chips/devices have been designed, fabricated, and on-line coupled with TRA-ICP-MS to achieve the goal. Moreover, the location and distribution of target analytes can be observed with the aid of laser ablation as sample introduction system for ICP. These methodologies have been employed for the investigation of endogenous elements/biomolecular in single cells and exogenous metal/nanoparticles/metal-containing drugs uptake by single cells.