Evaluation of Label-Free Confocal Raman Microspectroscopy for Monitoring Oxidative Stress In Vitro in Live Human Cancer Cells.
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Surmacki, J. M., Quiros-Gonzalez, I., & Bohndiek, S. E. (2022). Evaluation of Label-Free Confocal Raman Microspectroscopy for Monitoring Oxidative Stress In Vitro in Live Human Cancer Cells.. https://doi.org/10.3390/antiox11030573
Funder: Wellcome Trust
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 (<i>N</i>-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<sup>-1</sup> 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<sup>-1</sup>). 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.
Fluorescence microscopy, Oxidative stress, Raman microspectroscopy, tert-butyl hydroperoxide, N-acetyl-L-cysteine
Cancer Research UK (C14303/A17197, C47594/A16267)
European Union (FP7-PEOPLE-2013-CIG-630729)
External DOI: https://doi.org/10.3390/antiox11030573
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336410
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/