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

Antioxidants

Isabel Quiros-Gonzalez¹  Jakub Maciej Surmacki¹  Sarah Elizabeth Bohndiek¹ 

[1] Department of Physics, University of Cambridge, JJ Thomson Avenue, Cambridge CB3 0HE, UK;

关键词: oxidative stress;    Raman microspectroscopy;    fluorescence microscopy;    tert-butyl hydroperoxide;    N-acetyl-l-cysteine;   

DOI  :  10.3390/antiox11030573

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown