{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T17:22:02Z","timestamp":1793726522526,"version":"4.6.1"},"reference-count":51,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"27","content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2025,7,4]]},"abstract":"<jats:p>\n            Population heterogeneity in bacterial phenotypes, such as antibiotic resistance, is increasingly recognized as a medical concern. Heteroresistance occurs when a predominantly susceptible bacterial population harbors a rare resistant subpopulation. During antibiotic exposure, these resistant bacteria can be selected and lead to treatment failure. Standard antibiotic susceptibility testing methods often fail to reliably detect these subpopulations due to their low frequency, highlighting the need for improved diagnostic approaches. Here, we present a droplet microfluidics method where bacteria are encapsulated in droplets containing growth medium and antibiotics. The growth of rare resistant cells is detected by observing droplet shrinkage under microscopy. We validated this method for three clinically important antibiotics in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            isolates obtained from bloodstream infections and showed that it can detect resistant subpopulations as infrequent as 10\n            <jats:sup>\u22126<\/jats:sup>\n            using only 200 to 300 droplets. In addition, we designed a multiplex microfluidic chip to increase the throughput of the assay.\n          <\/jats:p>","DOI":"10.1126\/sciadv.adv4558","type":"journal-article","created":{"date-parts":[[2025,7,4]],"date-time":"2025-07-04T17:58:27Z","timestamp":1751651907000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":39,"title":["Droplet microfluidics\u2013based detection of rare antibiotic-resistant subpopulations in\n            <i>Escherichia coli<\/i>\n            from bloodstream infections"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0943-6751","authenticated-orcid":true,"given":"Sagar N.","family":"Agnihotri","sequence":"first","affiliation":[{"name":"Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden."},{"name":"Department of Materials Science and Engineering, Science for Life Laboratory, Uppsala University, Uppsala, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5081-0138","authenticated-orcid":true,"given":"Nikos","family":"Fatsis-Kavalopoulos","sequence":"additional","affiliation":[{"name":"Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2111-5291","authenticated-orcid":true,"given":"Jonas","family":"Windhager","sequence":"additional","affiliation":[{"name":"Department of Information Technology and SciLifeLab BioImage Informatics Facility, Uppsala University, Uppsala, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1264-1337","authenticated-orcid":true,"given":"Maria","family":"Tenje","sequence":"additional","affiliation":[{"name":"Department of Materials Science and Engineering, Science for Life Laboratory, Uppsala University, Uppsala, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6640-2174","authenticated-orcid":true,"given":"Dan I.","family":"Andersson","sequence":"additional","affiliation":[{"name":"Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","first-page":"277","article-title":"The antibiotic resistance crisis: Part 1: Causes and threats","volume":"40","author":"Ventola C. 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