Sensitive, high-throughput, metabolic analysis by molecular sensors on the membrane surface of mother yeast cells

Due to its genetic similarity to humans, yeast serves as a vital model organism in life sciences and medicine, allowing for the study of crucial biological processes such as cell division and metabolism for drug development. However, current tools for measuring yeast extracellular secretion lack the sensitivity, throughput, and speed required for large-scale metabolic analysis. Here, we present an ultrasensitive, large-scale analysis of yeast extracellular secretion using molecular sensors on the membrane surface of mother yeast cells. These sensors remain selectively confined to mother yeast cells during cell division, enabling high-sensitivity detection, high-throughput screening and rapid single-yeast assays. Their detection limit is 100 nM, and they can screen over 10⁷ single cells per run. We achieve a > 30-fold speed boost compared to conventional droplet-based screening, allowing us to identify the top 0.05% of secretory strains from 2.2 × 10⁶ variants within just 12 minutes. The platform offers potential for large-scale single-yeast metabolic analysis and bio-fabrication.