Fluorescence imaging-based methods for single-cell protein analysis

The quantity and activity of proteins in many biological systems exhibit prominent heterogeneities. Single-cell analytical methods can resolve subpopulations and dissect their unique signatures from heterogeneous samples, enabling a clarifying view of the biological process. Over the last 5 years, technologies for single-cell protein analysis have significantly advanced. In this article, we highlight a branch of those technology developments involving fluorescence-based approaches, with a focus on the methods that increase the ability to multiplex and enable dynamic measurements. We also analyze the limitations of these techniques and discuss current challenges in the field, with the hope that more transformative platforms can soon emerge.