Raman spectroscopic characterization of necrotic cell death.
Raman spectroscopy has been used to estimate the biochemical changes due to necrosis in an in vitro model system comprised of a human malignant melanoma cell line (MEL-28). Combined oxygen and glucose deprivation was used to simulate necrotic cell death in tumors. Raman spectroscopy measurements of nonproliferating live cells and dead cells were made at 24, 48, and 72 hours. Quantitative estimates of the biochemical composition of live and dead cells were made by fitting cell spectra to the basis spectra of protein, lipid, RNA, DNA, and glycogen. A decrease in the relative amount of lipid and RNA, and an increase in the relative protein content, were observed in dead cells. A comparison of the spectra indicated the existence of conformational changes in protein and nucleic acids in dead cells. These results suggest that Raman spectroscopy could be used to detect necrotic cell death in tumors.
Kunapareddy
Nagapratima
N
Los Alamos National Laboratory, Bioscience Division, MS E535, Los Alamos, New Mexico 87545, USA.
Freyer
James P
JP
Mourant
Judith R
JR
D017209:Apoptosis; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000032:analysis*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008055:Lipids / Q000032:analysis*; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology*; D009336:Necrosis; D009363:Neoplasm Proteins / Q000032:analysis*; D012334:RNA, Neoplasm / Q000032:analysis*; D013059:Spectrum Analysis, Raman / Q000295:instrumentation* / Q000379:methods*
D004273:DNA, Neoplasm; D008055:Lipids; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm