Abstract
Previous works showed that spatially resolved Raman spectra of cytoplasm and nucleus region of single cells exposed to X-rays evidence different features. The present work aims to introduce a new approach to profit from these differences to deeper investigate X-ray irradiation effects on single SH-SY5Y human neuroblastoma cells. For this aim, Raman micro-spectroscopy was performed in vitro on single cells after irradiation by graded X-ray doses (2, 4, 6, 8 Gy). Spectra from nucleus and cytoplasm regions were selectively acquired. The examination by interval Principal Component Analysis (i-PCA) of the difference spectra obtained by subtracting each cytoplasm-related spectrum from the corresponding one detected at the nucleus enabled us to reveal the subtle modifications of Raman features specific of different spatial cell regions. They were discussed in terms of effects induced by X-ray irradiation on DNA/RNA, lipids, and proteins. The proposed approach enabled us to evidence some features not outlined in previous investigations.
Highlights
The potentialities of Raman Spectroscopy (RS) as a diagnostic tool for biofluids, cells, and tissues have been widely demonstrated [1,2,3,4,5,6,7]
The average vector normalized spectra obtained for both nucleus and cytoplasm regions for control cells are shown in Figure 1 in the fingerprint region (400–1750 cm−1 ) and in Figure 2 in the high Raman shift region (HSR) (2650–3200 cm−1 )
The X-ray irradiation effects on the nuclear components of single SH-SY5Y human neuroblastoma cells irradiated by graded X-ray doses
Summary
The potentialities of Raman Spectroscopy (RS) as a diagnostic tool for biofluids, cells, and tissues have been widely demonstrated [1,2,3,4,5,6,7]. RS is a vibrational technique that enables to get information on the functional groups present in the sample under investigation, providing a molecular fingerprint of the sample itself. This technique attracts much interest because it allows one to yield this information in physiological conditions at various spatial scales, down to the single cell level or single cell region, thanks to the use of Raman micro-spectroscopy approach (μ-RS) [8,9,10]. Spectral difference and multivariate data analysis methods, such as Principal Component
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