Single-cell proteomics has seen significant advancements in recent years, enabling the analysis of thousands of proteins within single cells. These improvements are driven by advancements in experimental design, sample preparation, separation techniques, mass spectrometry instrumentation, and data acquisition/analysis. Label-free and multiplexed methods are both being actively developed, each with unique advantages. Advances in sample preparation have simplified workflows and optimized reagents, making the process more affordable and efficient. Ultra-low-flow nanoLC, microfabricated pillar array columns, and capillary electrophoresis have significantly improved sensitivity and resolution. The exploration of spatial proteomics with single-cell resolution holds promise, offering high-resolution tissue phenotyping. Applications of single-cell proteomics are expected to be highly impactful, particularly in cancer research, biomarker discovery, and developmental biology. Challenges remain, such as limited depth of proteome coverage per cell and the few cells that can be measured per day, but there are clear paths forward to improve these aspects. Single-cell proteomics stands at the threshold of uncovering novel frontiers in life sciences and addressing urgent challenges in medicine and biology.Single-cell proteomics has seen significant advancements in recent years, enabling the analysis of thousands of proteins within single cells. These improvements are driven by advancements in experimental design, sample preparation, separation techniques, mass spectrometry instrumentation, and data acquisition/analysis. Label-free and multiplexed methods are both being actively developed, each with unique advantages. Advances in sample preparation have simplified workflows and optimized reagents, making the process more affordable and efficient. Ultra-low-flow nanoLC, microfabricated pillar array columns, and capillary electrophoresis have significantly improved sensitivity and resolution. The exploration of spatial proteomics with single-cell resolution holds promise, offering high-resolution tissue phenotyping. Applications of single-cell proteomics are expected to be highly impactful, particularly in cancer research, biomarker discovery, and developmental biology. Challenges remain, such as limited depth of proteome coverage per cell and the few cells that can be measured per day, but there are clear paths forward to improve these aspects. Single-cell proteomics stands at the threshold of uncovering novel frontiers in life sciences and addressing urgent challenges in medicine and biology.