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Evaluation of Label-Free Confocal Raman Microspectroscopy for Monitoring Oxidative Stress In Vitro in Live Human Cancer Cells
| Content Provider | MDPI |
|---|---|
| Author | Surmacki, Jakub Maciej Quiros-Gonzalez, Isabel Bohndiek, Sarah Elizabeth |
| Copyright Year | 2022 |
| Description | 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. |
| Starting Page | 573 |
| e-ISSN | 20763921 |
| DOI | 10.3390/antiox11030573 |
| Journal | Antioxidants |
| Issue Number | 3 |
| Volume Number | 11 |
| Language | English |
| Publisher | MDPI |
| Publisher Date | 2022-03-17 |
| Access Restriction | Open |
| Subject Keyword | Antioxidants Analytical Chemistry Oxidative Stress Raman Microspectroscopy Fluorescence Microscopy Tert-butyl Hydroperoxide N-acetyl-l-cysteine |
| Content Type | Text |
| Resource Type | Article |
