Cell apoptosis specific marker found by Fourier Transform Infrared Spectroscopy

Abstract

We used Fourier Transform Infrared Spectroscopy (FTIR) combined with flow cytometry to study the apoptosis and necrosis processes in Jurkat, a lymphocyte cell line. The apoptosis was induced in the cells by a chemical agent, the actinomycin D, while the necrosis was induced lowering the pH value to 4.2. The apoptotic events were analysed by flow cytometry (using annexin V and propidium iodide) and contemporary monitored by FTIR spectroscopy at different times after the treatment. This comparison allowed us to find in the IR spectrum, between 3000 cm −1 and 2800 cm −1 , a "marker band" of the apoptosis corresponding to the exposure of phosphatidylserine on the outer leaflet of the membrane. A marker of a specific cellular process obtained by using a non-destructive technique such as FTIR spectroscopy, has a great significance in the diagnostic medicine providing a tool for detecting pathologies in vivo.

Analysis of postmortem metabolic changes in rat kidney cortex using Fourier transform infrared spectroscopy

“…2, there was a dramatic increase in this region whose spectrum is populated by absorptions stemming from the C-H stretching vibrations of olefinic =CH, -CH 2 and −CH 3 groups [18,19,23,25]. Band assignment (see [20][21][22][23][24][25][26][27][28][29]). As illustrated in Fig.…”

Growth substrate induced functional changes elucidated by FTIR and Raman spectroscopy in in–vitro cultured human keratinocytes

“…At present Fourier Transform Infrared Microspectrosopy (FTIRM) and Confocal Raman Microspectroscopy (CRM) have identified in-situ molecular alterations associated with cell death via apoptosis [1][2][3] or necrosis [4,5], or as a result of proliferative changes in cellular activity [6][7][8] or mitosis [9]. It has also been shown that these modalities can identify molecular changes associated with changes in the phenotype of differentiating embryonic cells [10,11].…”

Fourier Transform Infrared spectroscopy discloses different types of cell death in flow cytometrically sorted cells

“…Gaudenzi et al (2004) found a positive correlation between the vibrational band of ~2928 cm -1 increasing in intensity and the detection of the % apoptosis using annexin-V. Annexin-V binds to phosphatidylserine, which is a phospholipid externalized during apoptosis. It was speculated by Gaudenzi et al (2004) that the vibrational band could be directly attributed to phosphatidylserine (Gaudenzi et al, 2004).…”

“…FTIR-and Raman spectroscopy have demonstrated biomedical applications in cancer research (Diem et al, 2012;Mantsch et al, 2002) including the ability to differentiate between healthy and cancerous cells / tissues, spectral differences between different tissues and types of cancers, grading of the stage of cancer progression and drug discovery where new drug leads were tested against the proliferation of cancerous cells as well as investigations on the molecular mechanism of action of known chemotherapeutics. Some spectral regions have been identified where cell death induction led to spectral differences in FTIR data (Gaudenzi et al, 2004;Holman et al, 2000;Jamin et al, 2003;Munro et al, 2010;Notingher et al, 2003). In this investigation, we set out to determine whether FTIR microspectroscopy could successfully distinguish between viable, apoptotic and necrotic cells.…”