TY - JOUR A1 - Lipiec, E. A1 - Bambery, K. R. A1 - Heraud, P. A1 - Kwiatek, W.M. A1 - McNaughton, D. A1 - Tobin, M.J. A1 - Vogel, Christian A1 - Wood, B. R. T1 - Monitoring UVR induced damage in single cells and isolated nuclei using SR-FTIR microspectroscopy and 3D confocal Raman imaging N2 - SR-FTIR in combination with Principal Component Analysis (PCA) was applied to investigate macromolecular changes in a population of melanocytes and their extracted nuclei induced by environmentally relevant fluxes of UVR (Ultraviolet Radiation). Living cells and isolated cellular nuclei were investigated post-irradiation for three different irradiation dosages (130, 1505, 15 052 Jm-2 UVR, weighted) after either 24 or 48 hours of incubation. DNA conformational changes were observed in cells exposed to an artificial UVR solar-simulator source as evidenced by a shift in the DNA asymmetric phosphodiester vibration from 1236 cm-1 to 1242 cm-1 in the case of the exposed cells and from 1225 cm-1 to 1242 cm-1 for irradiated nuclei. PCA Scores plots revealed distinct clustering of spectra from irradiated cells and nuclei from non-irradiated controls in response to the range of applied UVR radiation doses. 3D Raman confocal imaging in combination with k-means cluster analysis was applied to study the effect of the UVR radiation exposure on cellular nuclei. Chemical changes associated with apoptosis were detected and included intra-nuclear lipid deposition along with chromatin condensation. The results reported here demonstrate the utility of SR-FTIR and Raman spectroscopy to probe in situ DNA damage in cell nuclei resulting from UVR exposure. These results are in agreement with the increasing body of evidence that lipid accumulation is a characteristic of aggressive cancer cells, and are involved in the production of membranes for rapid cell proliferation. PY - 2014 U6 - https://doi.org/10.1039/c4an00838c SN - 0003-2654 SN - 1364-5528 VL - 139 IS - 17 SP - 4200 EP - 4209 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-31172 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Lipiec, E. A1 - Bambery, K. R. A1 - Heraud, P. A1 - Hirschmugl, C. A1 - Lekki, J. A1 - Kwiatek, W.M. A1 - Tobin, M.J. A1 - Vogel, Christian A1 - Whelan, D. A1 - Wood, B. R. T1 - Synchrotron FTIR shows evidence of DNA damage and lipid accumulation in prostate adenocarcinoma PC-3 cells following proton irradiation N2 - Synchrotron Radiation Fourier Transform Infrared (SR-FTIR) spectra of single human prostate adenocarcinoma PC-3 cells, irradiated with a defined number of 2 MeV protons generated by a proton microbeam along with non-irradiated control cells, were analysed using multivariate methods. A number of different Principal Component Analysis (PCA) models were tested and the spectral ranges associated with nucleic acids, proteins and lipids were analysed separately. The results show a dose dependent shift of the O—P—O asymmetric stretching mode from 1234 cm-1 to 1237 cm-1, consistent with local disorder in the B-DNA conformation along with a change in intensity of the O—P—O symmetric stretching band at 1083 cm-1 indicative of chromatin fragmentation – the natural consequence of a high number of DNA Double Strand Breaks (DSBs). 2D mapping of characteristic functional groups at the diffraction limit shows evidence of lipid deposition and chromatin condensation in cells exposed to protons indicative of cell apoptosis following irradiation. These studies lay the foundation for understanding the macromolecular changes that occur to cells in response to radiation therapy, which has important implications in the treatment of tumours. KW - Synchrotron FTIR KW - Single cell KW - Radiation damage KW - Proton microbeam KW - DNA damage KW - Apoptosis PY - 2014 U6 - https://doi.org/10.1016/j.molstruc.2014.04.056 SN - 0022-2860 SN - 1872-8014 SN - 0377-046X VL - 1073 SP - 134 EP - 141 PB - Elsevier CY - Amsterdam AN - OPUS4-31173 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Lipiec, E. A1 - Bambery, K. R. A1 - Lekki, J. A1 - Tobin, M.J. A1 - Vogel, Christian A1 - Whelan, D. R. A1 - Wood, B. R. A1 - Kwiatek, W.M. T1 - SR-FTIR Coupled with principal component analysis shows evidence for the cellular bystander effect N2 - Synchrotron radiation-Fourier transform infrared (SR-FTIR) microscopy coupled with multivariate data analysis was used as an independent modality to monitor the cellular bystander effect. Single, living prostate cancer PC-3 cells were irradiated with various numbers of protons, ranging from 50–2,000, with an energy of either 1 or 2 MeV using a proton microprobe. SR-FTIR spectra of cells, fixed after exposure to protons and nonirradiated neighboring cells (bystander cells), were recorded. Spectral differences were observed in both the directly targeted and bystander cells and included changes in the DNA backbone and nucleic bases, along with changes in the protein secondary structure. Principal component analysis (PCA) was used to investigate the variance in the entire data set. The percentage of bystander cells relative to the applied number of protons with two different energies was calculated. Of all the applied quantities, the dose of 400 protons at 2 MeV was found to be the most effective for causing significant macromolecular perturbation in bystander PC-3 cells. PY - 2015 U6 - https://doi.org/10.1667/RR13798.1 SN - 0033-7587 SN - 1938-5404 VL - 184 IS - 1 SP - 73 EP - 82 PB - Radiation Research Society CY - Great Falls, Va. AN - OPUS4-33798 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -