Spectroscopic evaluation of living murine macrophage cells before and after activation using attenuated total reflectance infrared spectroscopy |
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| Affiliation: | 2. Department of Pathology, University of New Mexico Health Sciences Center, 915 Camino de Salud NE, Albuquerque, NM 87131, USA;1. Faculty of Foundry Engineering, AGH University of Science and Technology, 30-059 Krakow, Poland;2. Academic Centre for Materials and Nanotechnology, AGH University of Science and Technology, 30-059 Krakow, Poland;3. Department of Chemistry, Hankuk University of Foreign Studies, Yongin, Kyunggi-Do 449-791, Republic of Korea;1. Department of Neurobiology, Institute of Pharmacology, Polish Academy of Sciences, Smętna 12, 31-343 Kraków, Poland;2. Faculty of Foundry Engineering, AGH University of Science and Technology, ul. Reymonta 23, 30-059 Kraków, Poland;3. Institute of Nuclear Physics, Polish Academy of Science, 31-342 Krakow, Poland;4. Faculty of Chemistry, Jagiellonian University, ul. Ingardena 3, 30-060 Kraków, Poland;5. Department of Chemistry, Hankuk University of Foreign Studies, Yongin, Kyunggi-Do, 449-791, South Korea;1. Dipartimento di Scienze Fisiche e Chimiche, Università dell’Aquila, Via Vetoio 2, 67100, Coppito, L’Aquila, Italy;2. Sorbonne Universities, UPMC, Laboratoire de Réactivité de Surface UMR CNRS 7197, Tour 43-53, 3rd floor, 4 Pl. Jussieu, 75005, Paris, France;3. Service de Bioénergétique, Biologie Structurale et Mécanismes (SB2SM) CEA, iBiTec-S, Biochimie Biophysique et Biologie Structurale (B3S), I2BC, UMR 9198, F-91191 Gif-sur-Yvette, France;1. Uzhhorod National University, 46 Pidhirna Str., 88000, Uzhhorod, Ukraine;2. Vlokh Institute of Physical Optics, 23 Dragomanov Str., 79005, Lviv, Ukraine;3. Semiconductor Physics, Chemnitz University of Technology, D-09107, Chemnitz, Germany;4. Institute of Electron Physics, Ukr. Nat. Acad. Sci., 21 Universytetska Str., 88017, Uzhhorod, Ukraine |
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| Abstract: | Infection activates immune response pathways in host macrophages and lymphocytes that might be of sufficient magnitude to facilitate early diagnoses of infections through a host immune biosignature. Attenuated total reflectance (ATR) infrared spectroscopy was used to examine the spectroscopic signatures of living mouse macrophage cells before and after activation. Cells were prepared as control samples, or activated with a combination of lipopolysaccharide and interferon-gamma (IFN-γ) and analyzed 21 h after treatment. Resulting ATR/IR spectra collected from the living cells were analyzed using principal components analysis (PCA) and other classification methods. Plotting the scores from the first two principal components against one another provides good separation between activated and control samples. Interpretation of the loadings plots indicated that cellular activation was associated with changes in nucleic acid, protein and lipid infrared bands. Spectral samples were used to develop classification models based on activation status. Linear discriminant analysis (LDA) and K-nearest neighbor (K-NN) models were developed with 100% classification rates, using leave-one-out cross-validation procedures. Activated macrophages can be distinguished from macrophages in the resting state by their ATR spectroscopy biosignatures. |
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