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Recent advances discussed include high-throughput access to microbial dark matter, activation of cryptic BGCs via microfluidic co-culture, faster chemical dereplication with mass spectrometry integration, and linking resistance genes to individual hosts and single-cell 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bottlenecks in antibiotics discovery  \nHassan Barakat,1,2 Makenzie Bolton,1,2 Jingjing Cheng,1 and Xiangpeng Li1,3,*  \n1Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA  \n2These authors contributed equally  \n3Senior author  \n*[Correspondence:](Correspondence: xli@chem.fsu.edu)[ xli@chem.fsu.edu](Correspondence: xli@chem.fsu.edu)  \n[https://doi.org/10.1016/j.isci.2025.114377](https://doi.org/10.1016/j.isci.2025.114377)  \nSUMMARY  \nAntimicrobial resistance (AMR) presents a mounting global health crisis. Traditional antibiotic discovery methods are hindered by low throughput, frequent rediscovery of known compounds, and limited access to unculturable microbes and silent biosynthetic gene clusters (BGCs) . Droplet microfluidics offers a transformative platform to overcome these barriers by enabling massively parallel, compartmentalized analysis atthe single-cell level. In this perspective, we discuss how droplet microfluidics is reshaping the antibiotic discovery landscape by addressing long-standing limitations in cultivation, screening, and compound identification. We highlight recent advances in: (1) accessing microbial dark matter through high-throughput single-cell technologies,(2) activating cryptic BGCs and dissecting microbial interactions using microfluidiccoculture platforms,(3) accelerating chemical dereplication and discovery of novel metabolites through integration with mass spectrometry-based tools, and (4) uncovering resistance and persistence mechanisms by linking antibiotic resistance genes to individual microbial hosts and single cell transcriptomes. Together, these innovations position droplet microfluidics as a powerful engine to accelerate antibiotic discovery and combat the global threat of AMR.  \nINTRODUCTION  \nThe discovery of antibiotics revolutionized modern medicine by saving millions of lives, alleviating suffering, and transforming once-deadly infections into manageable conditions. However, the widespread and often indiscriminate use of antibiotics has driven the emergence of antimicrobial resistance (AMR) .1 ,2 In 2021, bacterial AMR was responsible for an estimated 5 million deaths, and this number is projected to increase substantially by 2050.3 If left unchecked, the rise of AMR could usher in a‘‘post-antibiotic era,’’ where simple infections and routine surgeries once again become life-threatening.3 ,4 To prevent this scenario, there is an urgent need to accelerate the discovery of new antibiotics.  \nRESULTS  \nThe major challenge in antibiotic discovery: Limited screening throughput  \nDespite the urgent need for new antibiotics, the pace of discovery has slowed dramatically over the past several decades.5 Classical antibiotic discovery—pioneered by Waksman in the 1940s—was based on isolating soil-derived microbes and screening them for antimicrobial activity.6 This approach yielded many antibiotics still in use today. But by the 1970s, much of the easily accessible chemical space had been explored, and the  \nmomentum stalled.5 This decline reflects multiple barriers, including limited financial incentives, the complexities of antibiotic stewardship, and regulatory challenges.7 ,8 Yet at the core of the scientific slowdown lies a fundamental bottleneck: low screening throughput (Box 1) . To accelerate antimicrobial drug development, higher-throughput screening technologies are urgently needed.  \nBreaking the antibiotic discovery bottleneck with droplet microfluidics  \nDroplet-based microfluidics involves the generation, manipulation, and control of sub-microliter droplets within an immiscible carrier fluid.9 By leveraging hydrodynamic forces and interfacial tension, this technology transforms continuous fluid streams into discrete, compartmentalized reaction volumes. Since the first demonstration of droplet generations in microfluidic systems in 1997,10 the field","cbCaifXjJR2SP6nG","https://ap.wps.com/l/cbCaifXjJR2SP6nG","pdf",2259575,12,"English","# Summary\n# Introduction\n# Results\n## The major challenge in antibiotic discovery: Limited screening throughput\n## Breaking the antibiotic discovery bottleneck with droplet microfluidics","[{\"question\":\"What major bottleneck slows antibiotic discovery, as described in the document?\",\"answer\":\"The document identifies low screening throughput as a fundamental bottleneck that limits acceleration of antimicrobial drug development.\"},{\"question\":\"How does droplet microfluidics help address antibiotic discovery limitations?\",\"answer\":\"Droplet microfluidics enables massively parallel, compartmentalized analysis at the single-cell level, supporting improved cultivation, screening, and compound identification.\"},{\"question\":\"Which four recent advances does the perspective highlight?\",\"answer\":\"It highlights access to microbial dark matter, activation of cryptic biosynthetic gene clusters and analysis via microfluidic co-culture, chemical dereplication and novel metabolite discovery with mass spectrometry integration, and linking resistance/persistence mechanisms to antibiotic resistance genes in individual hosts and single-cell transcriptomes.\"}]","Droplet microfluidics strategies for overcoming bottlenecks in antibiotics discovery | PDF",1790734211]