{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T18:29:29Z","timestamp":1766082569838,"version":"3.48.0"},"reference-count":75,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie","award":["713669"],"award-info":[{"award-number":["713669"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Stress tolerant slow- or non-growing bacterial subpopulations represent a key factor for chronic, recurrent, antibiotic-tolerant infections and necessitate prolonged antibiotic therapy. Consequently, new tools that facilitate investigation of these small subpopulations are highly needed. To characterize slow-\/non-growing populations of mycobacteria, we here utilized Vybrant DiD, a lipophilic, fluorescent organic probe that detected pathogenic and non-pathogenic species in a rapid, specific and non-invasive manner. It enabled accurate, direct quantification of mycobacterial replication rate\n                    <jats:italic>in vitro<\/jats:italic>\n                    and in murine macrophages at the population and single-cell level, providing insight into population heterogeneity. We determined the preexisting slow-\/non-growing fraction of mycobacteria during exponential growth, which increased upon stress. Monitoring mycobacterial resuscitation after antibacterial treatment and after low-oxygen-induced dormancy revealed the stochastic and heterogeneous nature of the resuscitation process. We anticipate that this method will be widely utilized in basic research on bacterial persistence and may also be included in applied settings, e.g., high-throughput drug characterization.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0338563","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T18:24:47Z","timestamp":1766082287000},"page":"e0338563","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid and efficient labeling by a selective organic fluorophore probe highlights heterogeneity of mycobacterial populations and persister resuscitation"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9910-4747","authenticated-orcid":true,"given":"Priyanka","family":"Chauhan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sander","family":"van Otterdijk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6991-3022","authenticated-orcid":true,"given":"Susanna","family":"Commandeur","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dirk","family":"Bald","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0255-4766","authenticated-orcid":true,"given":"Frank J.","family":"Bruggeman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"issue":"8","key":"pone.0338563.ref001","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1038\/nrmicro3491","article-title":"A functional perspective on phenotypic heterogeneity in microorganisms","volume":"13","author":"M Ackermann","year":"2015","journal-title":"Nat Rev Microbiol"},{"issue":"2","key":"pone.0338563.ref002","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1038\/nrmicro2504","article-title":"Microbial seed banks: the ecological and evolutionary implications of dormancy","volume":"9","author":"JT Lennon","year":"2011","journal-title":"Nat Rev Microbiol"},{"issue":"23","key":"pone.0338563.ref003","doi-asserted-by":"crossref","first-page":"4645","DOI":"10.1016\/j.jmb.2019.06.024","article-title":"Phenotypic diversification of microbial pathogens-cooperating and preparing for the future","volume":"431","author":"L Schr\u00f6ter","year":"2019","journal-title":"J Mol Biol"},{"issue":"9","key":"pone.0338563.ref004","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1038\/s41579-022-00721-0","article-title":"Types and functions of heterogeneity in mycobacteria","volume":"20","author":"ES Chung","year":"2022","journal-title":"Nat Rev Microbiol"},{"issue":"6","key":"pone.0338563.ref005","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1016\/j.chom.2013.05.009","article-title":"Microbial persistence and the road to drug resistance","volume":"13","author":"NR Cohen","year":"2013","journal-title":"Cell Host Microbe"},{"key":"pone.0338563.ref006","doi-asserted-by":"crossref","DOI":"10.1038\/emi.2014.3","article-title":"Persisters, persistent infections and the Yin-Yang model","volume-title":"Emerging Microbes and Infections","author":"Y Zhang","year":"2014"},{"key":"pone.0338563.ref007","unstructured":"WHO. 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