Single-cell proteomic analyses are of fundamental importance in order to capture biological heterogeneity within complex cell systems and heterogeneous populations. Herein, we developed an automated and multiplexed sample processing workflow combining integrated stationary nanoliter-droplet chips and a commercial liquid dispensing instrument for single-cell proteomics. Utilizing this robust workflow and data independent acquisition (DIA) mode, we can profile up to 3000 protein groups in single mammalian cell with a remarkable improvement, achieving a dynamic range of 6 orders of magnitude with good reproducibility. The chip-based system was further applied in single lymphocyte analysis, with protein groups of 617 and 1089 identified at the single B and T lymphocyte, demonstrating the promising applications of single-cell insights in clinical research, especially in cancer immunology.