Abstract
FT-IR is used in the field of biology and medicine to detect bimolecular changes in disordered cells and tissues. In this report, using FT-IR microscopy, we characterize changes in apoptotic and necrotic Jurkat cells with respect to normal cells. The analysis of deconvoluted regions of the FT-IR spectra showed significant differences compared to the controls in three spectral regions. In particular, the apoptotic cells were characterized by an increase in the absorption at 2925 cm−1, due to the asymmetric CH2-stretching (νasCH2) of membrane lipids whereas the spectral areas ratio (A1654/A1629) of the amide I region indicated an increase in apoptotic cells of more α-helical structures with respect to ofβ-sheet content. Interestingly, apoptotic cells showed the appearance of a peak around 1743 cm¯1,ν(C=O) assigned to acid ester. Because no other similar increase for lipid bands was observed, the increase of A1745is not simply due to an increase in the number of lipid molecules or their density but could also be indicative as marker of apoptosis. These spectral changes were not observed in necrotic Jurkat cells.
Highlights
Techniques providing chemical information within living cells and tissues represent a very powerful tool for biomedical research directed toward the understanding of the causes of disease onset and on the properties and functions of diseased cells
Jurkat cells collected at different apoptotic stage or different necrotic stage were analyzed by FT-IR microscopy
We found that the peak area ratio A1558/A1540 slightly increased after treatment of the cells with drugs (0.58 ± 0.03 for normal cell and 0.58 ± 0.03, 0.68 ± 0.03 and 0.69 ± 0.03 for cell treated with VP-16 at 8, 16 and 32 h, or 0.62 ± 0.03 for cell treated with Cis-P at 8 and 16 h, respectively), suggesting an increase in the absorption of α-helix
Summary
Techniques providing chemical information within living cells and tissues represent a very powerful tool for biomedical research directed toward the understanding of the causes of disease onset and on the properties and functions of diseased cells. Like nucleic acids [35], proteins [34] and lipids [19] have characteristic and well-defined IR-active vibrational modes the analysis of IR spectra can detect, identify and quantify several molecular species within a biological sample. Under this aspects, FT-IR spectroscopy has been use to study variation in the conformation of biological macromolecules occurring between normal cells and cancer cells [1,2,4, 11,23,28,31]. FT-IR spectroscopy represents an important tool for the comprehension
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