title: Cell death stages in single apoptotic and necrotic cells monitored by Raman microspectroscopy.
abstract: Although apoptosis and necrosis have distinct features, the identification and discrimination of apoptotic and necrotic cell death in vitro is challenging. Immunocytological and biochemical assays represent the current gold standard for monitoring cell death pathways; however, these standard assays are invasive, render large numbers of cells and impede continuous monitoring experiments. In this study, both room temperature (RT)-induced apoptosis and heat-triggered necrosis were analyzed in individual Saos-2 and SW-1353 cells by utilizing Raman microspectroscopy. A targeted analysis of defined cell death modalities, including early and late apoptosis as well as necrosis, was facilitated based on the combination of Raman spectroscopy with fluorescence microscopy. Spectral shifts were identified in the two cell lines that reflect biochemical changes specific for either RT-induced apoptosis or heat-mediated necrosis. A supervised classification model specified apoptotic and necrotic cell death based on single cell Raman spectra. To conclude, Raman spectroscopy allows a non-invasive, continuous monitoring of cell death, which may help shedding new light on complex pathophysiological or drug-induced cell death processes.
authors.affiliation: 1] Fraunhofer-Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany [2] University of Stuttgart, Institute of Interfacial Process Engineering and Plasma Technology (IGVP), Nobelstr. 12, 70569 Stuttgart, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Brauchle
authors.affiliation: Fraunhofer-Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Thude
authors.affiliation: University Women's Hospital, Eberhard-Karls-University Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.
authors.forename: Sara Y
authors.initials: SY
authors.lastname: Brucker
authors.affiliation: 1] Fraunhofer-Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany [2] University Women's Hospital, Eberhard-Karls-University Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany [3] Department of Medicine/Cardiology, Cardiovascular Research Laboratories, David Geffen School of Medicine at UCLA, 675 Charles E. Young Drive South, MRL 3645, Los Angeles, CA, USA.
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000096:biosynthesis; D053178:Caspase 6 / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000473:pathology; D006358:Hot Temperature; D006801:Humans; D008856:Microscopy, Fluorescence; D009336:Necrosis / Q000503:physiopathology*; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D053148:Caspase 3; D053178:Caspase 6
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