{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T12:21:17Z","timestamp":1778070077738,"version":"3.51.4"},"reference-count":36,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2018,5,18]],"date-time":"2018-05-18T00:00:00Z","timestamp":1526601600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["GA16-12477S"],"award-info":[{"award-number":["GA16-12477S"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","award":["LO1212"],"award-info":[{"award-number":["LO1212"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","award":["CZ.1.05\/2.1.00\/01.0017"],"award-info":[{"award-number":["CZ.1.05\/2.1.00\/01.0017"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004240","name":"Akademie V\u011bd \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["RVO:68081731"],"award-info":[{"award-number":["RVO:68081731"]}],"id":[{"id":"10.13039\/501100004240","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["FKZ 01EO1502"],"award-info":[{"award-number":["FKZ 01EO1502"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["FKZ 13GW0096F"],"award-info":[{"award-number":["FKZ 13GW0096F"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"name":"COST Raman4Clinics","award":["BM 1401"],"award-info":[{"award-number":["BM 1401"]}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["JBIL"],"award-info":[{"award-number":["JBIL"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["FKZ PO 633\/29-1"],"award-info":[{"award-number":["FKZ PO 633\/29-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["BA 1601\/10-1"],"award-info":[{"award-number":["BA 1601\/10-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We tested optical micromanipulation in a microfluidic chamber chip by transferring individual bacteria into the chambers. We then subjected the cells in the chambers to antibiotic cefotaxime and we observed the changes by using time-lapse microscopy. Separately, we used laser tweezers Raman spectroscopy (LTRS) in a different micro-chamber chip to manipulate and analyze individual cefotaxime-treated E. coli cells. Additionally, we performed conventional Raman micro-spectroscopic measurements of E. coli cells in a micro-chamber. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. The principal component analysis (PCA) of Raman data distinguished between the cefotaxime treated cells and control. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments.<\/jats:p>","DOI":"10.3390\/s18051623","type":"journal-article","created":{"date-parts":[[2018,5,21]],"date-time":"2018-05-21T04:07:30Z","timestamp":1526875650000},"page":"1623","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress"],"prefix":"10.3390","volume":"18","author":[{"given":"Zden\u011bk","family":"Pil\u00e1t","sequence":"first","affiliation":[{"name":"Institute of Scientific Instruments (ISI), Czech Academy of Sciences, v.v.i., Kralovopolska 147, 612 64 Brno, Czech Republic"}]},{"given":"Silvie","family":"Bernatov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Scientific Instruments (ISI), Czech Academy of Sciences, v.v.i., Kralovopolska 147, 612 64 Brno, Czech Republic"}]},{"given":"Jan","family":"Je\u017eek","sequence":"additional","affiliation":[{"name":"Institute of Scientific Instruments (ISI), Czech Academy of Sciences, v.v.i., Kralovopolska 147, 612 64 Brno, Czech Republic"}]},{"given":"Johanna","family":"Kirchhoff","sequence":"additional","affiliation":[{"name":"Center for Sepsis Control and Care (CSCC), Jena University Hospital, Am Klinikum 1, D-07747 Jena, Germany"},{"name":"Institute of Physical Chemistry, Friedrich Schiller University Jena, Helmholtzweg 4, D-07743 Jena, Germany"},{"name":"Leibniz Institute of Photonic Technology (Leibniz IPHT), Albert-Einstein-Str. 9, D-07745 Jena, Germany"}]},{"given":"Astrid","family":"Tannert","sequence":"additional","affiliation":[{"name":"Center for Sepsis Control and Care (CSCC), Jena University Hospital, Am Klinikum 1, D-07747 Jena, Germany"},{"name":"Leibniz Institute of Photonic Technology (Leibniz IPHT), Albert-Einstein-Str. 9, D-07745 Jena, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7593-6132","authenticated-orcid":false,"given":"Ute","family":"Neugebauer","sequence":"additional","affiliation":[{"name":"Center for Sepsis Control and Care (CSCC), Jena University Hospital, Am Klinikum 1, D-07747 Jena, Germany"},{"name":"Institute of Physical Chemistry, Friedrich Schiller University Jena, Helmholtzweg 4, D-07743 Jena, Germany"},{"name":"Leibniz Institute of Photonic Technology (Leibniz IPHT), Albert-Einstein-Str. 9, D-07745 Jena, Germany"}]},{"given":"Ota","family":"Samek","sequence":"additional","affiliation":[{"name":"Institute of Scientific Instruments (ISI), Czech Academy of Sciences, v.v.i., Kralovopolska 147, 612 64 Brno, Czech Republic"}]},{"given":"Pavel","family":"Zem\u00e1nek","sequence":"additional","affiliation":[{"name":"Institute of Scientific Instruments (ISI), Czech Academy of Sciences, v.v.i., Kralovopolska 147, 612 64 Brno, Czech Republic"}]}],"member":"1968","published-online":{"date-parts":[[2018,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1038\/srep01258","article-title":"Integrated microfluidic device for single-cell trapping and spectroscopy","volume":"3","author":"Liberale","year":"2013","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"19021","DOI":"10.3390\/s150819021","article-title":"Review: Raman spectroscopy of optically trapped single biological micro-particles","volume":"15","author":"Redding","year":"2015","journal-title":"Sensors"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1002\/mrd.22284","article-title":"Metabolomics fingerprinting of the human seminal plasma of asthenozoospermic patients","volume":"81","author":"Gilany","year":"2014","journal-title":"Mol. Reprod. Dev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1364\/AO.32.000925","article-title":"Analysis of metabolites in aqueous solutions by using laser Raman spectroscopy","volume":"32","author":"Wang","year":"1993","journal-title":"Appl. Opt."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1626","DOI":"10.1128\/AEM.71.3.1626-1637.2005","article-title":"Chemotaxonomic identification of single bacteria by micro-Raman spectroscopy: Application to clean-room-relevant biological contaminations","volume":"71","author":"Harz","year":"2005","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1128\/JCM.01900-08","article-title":"Optical fingerprinting in bacterial epidemiology: Raman spectroscopy as a real-time typing method","volume":"47","author":"Jachtenberg","year":"2009","journal-title":"Clin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1002\/jrs.4844","article-title":"The application of Raman spectroscopy for the detection and identification of microorganisms","volume":"47","author":"Kirchhoff","year":"2016","journal-title":"J. Raman Spectrosc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6208","DOI":"10.1364\/OPEX.12.006208","article-title":"Real-time Raman spectroscopy of optically trapped living cells and organelles","volume":"25","author":"Xie","year":"2004","journal-title":"Opt. Express"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1800","DOI":"10.1364\/OL.30.001800","article-title":"Raman sorting and identification of single living micro-organisms with optical tweezers","volume":"30","author":"Xie","year":"2005","journal-title":"Opt. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2856","DOI":"10.1016\/S0006-3495(99)77117-1","article-title":"Characterization of photodamage to Escherichia coli in optical traps","volume":"77","author":"Neuman","year":"1999","journal-title":"Biophys. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1002\/cyto.a.20407","article-title":"Monitoring dynamic protein expression in living E. coli bacterial cells by laser tweezers Raman spectroscopy","volume":"71A","author":"Chan","year":"2007","journal-title":"Cytom. Part A"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Dai, J., Hamon, M., and Jambovane, S. (2016). Microfluidics for antibiotic susceptibility and toxicity testing. Bioengineering, 3.","DOI":"10.3390\/bioengineering3040025"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8343","DOI":"10.1007\/s00216-015-8912-y","article-title":"Identification of vancomycin interaction with Enterococcus faecalis within 30 min of interaction time using Raman spectroscopy","volume":"407","author":"Assmann","year":"2015","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1002\/jbio.201600316","article-title":"On-Chip spectroscopic assessment of microbial susceptibility to antibiotics within 3\u00bd hours","volume":"10","author":"Kirchhoff","year":"2017","journal-title":"J. Biophotonic"},{"key":"ref_15","first-page":"8271","article-title":"Detection of vancomycin resistances in enterococci within 3\u00bd hours","volume":"5","author":"Beleites","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1811","DOI":"10.1021\/acs.analchem.7b03800","article-title":"Simple ciprofloxacin resistance test and determination of minimal inhibitory concentration (MIC) within two hours using Raman spectroscopy","volume":"90","author":"Kirchhoff","year":"2018","journal-title":"Anal. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"10717","DOI":"10.1021\/ac4021616","article-title":"Combined dielectrophoresis-Raman setup for the classification of pathogens recovered from the urinary tract","volume":"85","author":"Ramoji","year":"2013","journal-title":"Anal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"044118","DOI":"10.1063\/1.4928070","article-title":"Rapid, culture-independent, optical diagnostics of centrifugally captured bacteria from urine samples","volume":"9","author":"Bokeloh","year":"2015","journal-title":"Biomicrofluidics"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Pil\u00e1t, Z., Jon\u00e1\u0161, A., Je\u017eek, J., and Zem\u00e1nek, P. (2017). Effects of infrared optical trapping on Saccharomyces cerevisiae in a microfluidic system. Sensors, 17.","DOI":"10.3390\/s17112640"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1364\/JOSAB.34.001855","article-title":"Thermal tuning of spectral emission from optically trapped liquid-crystal droplet resonators","volume":"34","author":"Aas","year":"2017","journal-title":"JOSA B"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"8635","DOI":"10.3390\/s100908635","article-title":"Raman Microspectroscopy of individual algal Cells: Sensing unsaturation of storage lipids in vivo","volume":"10","author":"Samek","year":"2010","journal-title":"Sensors"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"83060L","DOI":"10.1117\/12.912264","article-title":"Raman microspectroscopy of algal lipid Bodies: Beta-carotene as a sensor","volume":"8306","author":"Samek","year":"2011","journal-title":"SPIE Proc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1007\/s10811-011-9754-4","article-title":"Raman microspectroscopy of algal lipid Bodies: Beta-carotene quantification","volume":"24","author":"Samek","year":"2012","journal-title":"J. Appl. Phycol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"13188","DOI":"10.3390\/molecules181113188","article-title":"Following the mechanisms of bacteriostatic versus bactericidal action using Raman spectroscopy","volume":"18","author":"Samek","year":"2013","journal-title":"Molecules"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"23924","DOI":"10.3390\/ijms151223924","article-title":"Candida parapsilosis biofilm identification by Raman spectroscopy","volume":"15","author":"Samek","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"051038","DOI":"10.1117\/1.JBO.20.5.051038","article-title":"Identification of individual biofilm-forming bacterial cells using Raman tweezers","volume":"20","author":"Samek","year":"2015","journal-title":"J. Biomed. Opt."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"29635","DOI":"10.3390\/s151129635","article-title":"Influence of culture media on microbial fingerprints using Raman spectroscopy","volume":"15","author":"Samek","year":"2015","journal-title":"Sensors"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1088\/1464-4258\/9\/8\/S06","article-title":"Raman spectroscopy of optically trapped particles","volume":"9","author":"Petrov","year":"2007","journal-title":"J. Opt. A Pure Appl. Opt."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4813","DOI":"10.1002\/elps.200800484","article-title":"Light at work: The use of optical forces for particle manipulation, sorting, and analysis","volume":"29","year":"2008","journal-title":"Electrophoresis"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1146\/annurev.matsci.28.1.153","article-title":"Soft lithography","volume":"28","author":"Xia","year":"1998","journal-title":"Ann. Rev. Mater. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jphotobiol.2013.02.006","article-title":"Optical trapping of microalgae at 735\u20131064 nm: Photodamage assessment","volume":"121","author":"Nedbal","year":"2013","journal-title":"J. Photochem. Photobiol. B Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1366\/000370206776342553","article-title":"Optimization of the Rolling-Circle Filter for Raman Background Subtraction","volume":"60","author":"Brandt","year":"2006","journal-title":"Appl. Spectrosc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"267ra174","DOI":"10.1126\/scitranslmed.3009650","article-title":"A rapid antimicrobial susceptibility test based on single-cell morphological analysis","volume":"6","author":"Choi","year":"2014","journal-title":"Sci. Transl. Med."},{"key":"ref_34","unstructured":"Long, D.A. (2002). The Raman Effect: A Unified Treatment of the Theory of Raman Scattering by Molecules, John Wiley & Sons Ltd.. ISBNs 0-471-49028-8 (Hardback), 0-470-84576-7 (Electronic)."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1039\/b507715j","article-title":"Micro-Raman spectroscopic identification of bacterial cells of the genus Staphylococcus and dependence on their cultivation conditions","volume":"130","author":"Harz","year":"2005","journal-title":"Analyst"},{"key":"ref_36","first-page":"83","article-title":"Surface enhanced Raman spectra of Escherichia coli cells using ZnO nanoparticles","volume":"4","author":"Dutta","year":"2009","journal-title":"Dig. J. Nanomater. Biostruct."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/5\/1623\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:04:59Z","timestamp":1760195099000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/5\/1623"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,5,18]]},"references-count":36,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2018,5]]}},"alternative-id":["s18051623"],"URL":"https:\/\/doi.org\/10.3390\/s18051623","relation":{"has-preprint":[{"id-type":"doi","id":"10.20944\/preprints201804.0163.v1","asserted-by":"object"}]},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,5,18]]}}}