Silver nanoparticles (Ag NPs) are known to exhibit broad antimicrobial activity. However, such activity continues to raise concerns in the context of the interaction of such NPs with biomolecules. In a physiological environment NPs interact with individual biological cells either by penetrating through the cell membrane or by adhering to the membrane. We have explored the interaction of Ag NPs with single optically-trapped, live erythrocytes (red blood cells, RBCs) using Raman Tweezers spectroscopy. Our experiments reveal that Ag NPs induce modifications within an RBC that appear to be irreversible. In particular we are able to identify that the heme conformation in an RBC transforms from the usual R-state (oxy-state) to the T-state (deoxy-state). We rationalize our observations by proposing a model for the nanoparticle cytotoxicity pathway when the NP size is larger than the membrane pore size. We propose that the interaction of Ag NPs with the cell surface induces damage brought about by alteration of intracellular pH caused by the blockage of the cell membrane transport.

Advancements in the field of nanotechnology have resulted in the emergence of a large variety of engineered nanomaterials for innumerable applications. Despite the ubiquitous use of nanomaterials in daily life, concerns regarding the potential toxicity and safety of these materials have also been raised. There is a high demand for assessing the unwanted effects of both gold and silver nanoparticles, which is increasingly being used in biomedical applications. This paper deals with the study of stress due to silver and gold nanoparticles of varying size on red blood cells (RBCs) using Raman tweezers spectroscopy. RBCs were incubated with nanoparticles of size in the 10−100 nm range with the same concentrations, and micro-Raman spectra were recorded by optically trapping the nanoparticle-treated live RBCs. Spectral modifications implicating hemoglobin deoxygenation were observed in all nanoparticle-treated RBCs. One of the probable reason for the deoxygenation trend can be the adhesion of nanoparticles onto the cell surface causing imbalance in cell functioning. Moreover, the higher spectral variations observed for silver nanoparticles indicate that oxidative stress is involved in cell damage. These mechanisms lead to the modification in the hemoglobin structure because of changes in the pH of cytoplasm, which can be detected using Raman spectroscopy.

Abstract. Single-cell micro-Raman spectroscopy has been applied to explore cell differentiation in single, live, and malignant cells from two tumor cell lines. The spectra of differentiated cells exhibit substantial enhancement primarily in the intensities of protein peaks with concomitant decrease in intensities of O─P─O asymmetric stretching peaks in DNA/RNA. Principal component analyses show that the spectral score of differentiated cells tends to asymptotically approach that of spectra obtained from normal neural stem cells/progenitors. This lends credence to the notion that the observed spectral changes are specific to differentiation, since upon differentiation, malignant cells become less malignant and tend toward benignity. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Eryptosis—the suicidal death of erythrocytes—is characterized by membrane blebbing and cell shrinkage. Eryptosis can be triggered by various xenobiotics such as carbon monoxide, lead, and amyloid, and by stressors such as oxidative stress, osmotic shock, and rapid alteration of ambient conditions. We have used Raman tweezers spectroscopy to study eryptosis in single, live cells and have attempted to explore the underlying mechanism, specifically to identify possible Raman signatures of eryptosis. Erythrocytes (red blood cells) were exposed to free radicals, silver nanoparticles, glucose, heat, and osmotic shock to induce eryptosis, and a comparison was made of their Raman spectra, which indicated that these conditions lead to a transition of haemoglobin from the R to the T state. Consequences of eryptosis include dehydration, cell shrinkage, and pH changes, which result in deoxygenation of haemoglobin. This, in turn, can be detected by monitoring the wavenumber shifts associated with Raman marker bands of R to T transitions. In addition, the principal component analysis results indicate differentiation among red blood cells undergone eryptosis due to different conditions.

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.

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