Evaluation of Label-Free Confocal Raman Microspectroscopy for Monitoring Oxidative Stress In Vitro in Live Human Cancer Cells

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cam.issuedOnline2022-03-17
dc.contributor.authorSurmacki, Jakub Maciej
dc.contributor.authorQuiros-Gonzalez, Isabel
dc.contributor.authorBohndiek, Sarah Elizabeth
dc.contributor.orcidSurmacki, Jakub Maciej [0000-0001-6654-0286]
dc.contributor.orcidBohndiek, Sarah Elizabeth [0000-0003-0371-8635]
dc.date.accessioned2022-03-20T03:30:16Z
dc.date.available2022-03-20T03:30:16Z
dc.date.issued2022-03-17
dc.date.updated2022-03-20T03:30:16Z
dc.description.abstract<jats:p>Understanding the impact of free radicals and antioxidants in cell biology is vital; however, noninvasive nonperturbative imaging of oxidative stress remains a challenge. Here, we evaluated the ability of label-free Raman spectroscopy to monitor redox biochemical changes in antioxidant (N-acetyl-l-cysteine, NAC) and pro-oxidant (tert-butyl hydroperoxide, TBHP) environments. Cellular changes were compared to fluorescence microscopy using CellROX Orange as a marker of oxidative stress. We also investigated the influence of cell media with and without serum. Incubation of cells with NAC increased the Raman signal at 498 cm−1 from S-S disulphide stretching mode, one of the most important redox-related sensors. Exposure of cells to TBHP resulted in decreased Raman spectral signals from DNA/proteins and lipids (at 784, 1094, 1003, 1606, 1658 and 718, 1264, 1301, 1440, 1746 cm−1). Using partial least squares–discriminant analysis, we showed that Raman spectroscopy can achieve sensitivity up to 96.7%, 94.8% and 91.6% for control, NAC and TBHP conditions, respectively, with specificity of up to 93.5, 90.1% and 87.9%. Our results indicate that Raman spectroscopy can directly measure the effect of NAC antioxidants and accurately characterize the intracellular conditions associated with TBHP-induced oxidative stress, including lipid peroxidation and DNA damage.</jats:p>
dc.identifier.doi10.17863/CAM.82667
dc.identifier.eissn2076-3921
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/335237
dc.languageen
dc.publisherMDPI AG
dc.publisher.urlhttp://dx.doi.org/10.3390/antiox11030573
dc.subjectoxidative stress
dc.subjectRaman microspectroscopy
dc.subjectfluorescence microscopy
dc.subjecttert-butyl hydroperoxide
dc.subjectN-acetyl-l-cysteine
dc.titleEvaluation of Label-Free Confocal Raman Microspectroscopy for Monitoring Oxidative Stress In Vitro in Live Human Cancer Cells
dc.typeArticle
dcterms.dateAccepted2022-03-14
prism.issueIdentifier3
prism.publicationNameAntioxidants
prism.volume11
pubs.funder-project-idCancer Research UK (C14303/A17197, C47594/A16267)
pubs.funder-project-idEuropean Union (FP7-PEOPLE-2013-CIG-630729)
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0/
rioxxterms.versionVoR
rioxxterms.versionofrecord10.3390/antiox11030573
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