Micro-Raman spectroscopy study of optically trapped erythrocytes in malaria, dengue and leptospirosis infections.

Malaria, dengue and leptospirosis are three tropical infectious diseases that present with severe hematological derangement causing significant morbidity and mortality, especially during the seasonal monsoons. During the course of these infectious diseases, circulating red blood cells are imperiled to the direct ill-effects of the infectious pathogen in the body as well as to the pro-inflammatory cytokines generated as a consequence of the infection. RBCs when exposed to such inflammatory and/or pathogenic milieu are susceptible to injuries such as RBC programmed eryptosis or RBC programmed necrosis. This research aimed to explore the Raman spectra of live red cells that were extracted from patients infected with malaria, dengue, and leptospirosis. Red cells were optically trapped and micro-Raman probed using a 785 nm Diode laser. RBCs from samples of all three diseases displayed Raman signatures that were significantly altered from the normal/healthy. Distinct spectral markers that were common across all the four groups were obtained from various standardized multivariate analytical methods. Following comprehensive examination of multiple studies, we propose these spectral wavenumbers as "Raman markers of RBC injury." Findings in our study display that anemia-triggering infections can inflict variations in the healthy status of red cells, easily identifiable by selectively analyzing specific Raman markers. Additionally, this study also highlights relevant statistical tools that can be utilized to study Raman spectral data from biological samples which could help identify the very significant Raman peaks from the spectral band. This approach of RBC analysis can foster a better understanding of red cell behavior and their alterations exhibited in health and disease.

Micro-Raman Spectroscopy Analysis of Optically Trapped Erythrocytes in Jaundice.

Derangements in bilirubin metabolism and/or dysfunctions in the hepato-biliary system lead to the unhealthy buildup of bilirubin in blood, resulting in jaundice. During the course of this disorder, circulating red cells are invariably subjected to toxic effects of serum bilirubin and an array of inflammatory compounds. This study aimed to investigate the vibrational spectroscopy of live red cells in jaundice using micro-Raman spectroscopy combined with optical-trap. Red cells from blood samples of healthy volunteers and patients with jaundice were optically immobilized and micro-Raman probed using a 785 nm diode laser. Raman signatures from red cells in jaundice exhibited significant variations from the normal and the spectral-markers were obtained from multivariate analytical methods. This research gives insightful views on how different pathologies can act as "stress-milieus" for red cells in circulation, possibly impeding their normal functions and also exasperating anemia. Raman spectroscopy, an emerging bio-analytical technique, is sensitive in detecting molecular-conformations in situ, at cellular-levels and in real-time. This study could pave way in understanding fundamental red cell behavior in different diseases by analyzing Raman markers.

Influence of static electric field on Raman polarizability of optically trapped polystyrene beads.

The influence of static electric field of about 4.9 kV/cm on optically trapped polystyrene bead has been studied using micro-Raman spectroscopy. This experimental method could explore the response of electronic polarizability of molecules to the static electric field by measuring the Raman intensity. It is observed that, the atomic electrons in polystyrene will polarize to the extent that they induce a Raman intensity enhancements by a factor ranging from 0.004 to 0.64 for various spectral bands of polystyrene when exposed to the electric field from ~4.90 to 4.96 kV/cm.

A micro-Raman and chemometric study of urinary tract infection-causing bacterial pathogens in mixed cultures.

Detection of urinary tract infection (UTI)-causing bacteria uses conventional time-consuming microbiological techniques. The current need is to use a fast and reliable method of bacterial identification. In order to unambiguously distinguish the UTI-causing five bacterial species used in the current study, micro-Raman spectra were obtained from a home-assembled micro-Raman system and analyzed by multivariate statistical techniques such as principal component analysis (PCA), partial least square-discriminate analysis (PLS-DA), and support vector machine (SVM). Also, the micro-Raman spectra recorded from samples containing two and three bacterial species were tested and validated against the aforementioned calibration models using PLS-DA and SVM. The prediction accuracies of up to 73 and 89% were achieved with PLS-DA and SVM, respectively. Taken together, the present study depicts the capturing of unique micro-Raman spectral features manifesting from the biochemical content of each bacterium. Also, micro-Raman spectroscopy combined with multivariate data analysis can therefore be a reliable and faster technique for the diagnosis of UTI-causing bacteria. Graphical Abstract.