Artificial Intelligence-Assisted Automatic Raman-Activated Cell Sorting (AI-RACS) System for Mining Specific...

The microbiome represents the natural presence of microorganisms, and exploring, understanding, and leveraging its functions will bring about significant breakthroughs in life sciences and applications. Raman-activated cell sorting (RACS) enables the correlation of phenotype and genotype at the single-cell level, offering a solution to the bottleneck in microbial community functional analysis caused by challenges in cultivating diverse microorganisms. However, current labor-intensive manual procedures fall short in catering to the demands of single-cell functional analysis in microbial communities. To address this issue, we developed an artificial intelligence-assisted Raman-activated cell sorting system (AI-RACS) that integrates precise single-cell positioning, automated data collection, optical tweezers capture, and single-cell printing to elevate microbial single-cell RACS from manual to automated, validating the efficacy of the system by isolating aluminum-tolerant microbes from acidic soil microbiota. Leveraging the AI-RACS framework, we sorted 13 strains from red soil samples under near-in situ conditions, with all demonstrating strong aluminum tolerance. AI-RACS efficiently segregates microbial cells from intricate environmental samples, investigating their functional attributes and presenting a novel tool for microbial research and applications.