Identification of Oxidative Stress in Red Blood Cells with Nanoscale Chemical Resolution by Infrared Nanospectroscopy

Abstract

During their lifespan, Red blood cells (RBC), due to their inability to self-replicate, undergo an ageing degradation phenomenon. This pathway, both in vitro and in vivo, consists of a series of chemical and morphological modifications, which include deviation from the biconcave cellular shape, oxidative stress, membrane peroxidation, lipid content decrease and uncoupling of the membrane-skeleton from the lipid bilayer. Here, we use the capabilities of atomic force microscopy based infrared nanospectroscopy (AFM-IR) to study and correlate, with nanoscale resolution, the morphological and chemical modifications that occur during the natural degradation of RBCs at the subcellular level. By using the tip of an AFM to detect the photothermal expansion of RBCs, it is possible to obtain nearly two orders of magnitude higher spatial resolution IR spectra, and absorbance images than can be obtained on diffraction-limited commercial Fourier-transform Infrared (FT-IR) microscopes. Using this approach, we demonstrate that we can identify localized sites of oxidative stress and membrane peroxidation on individual RBC, before the occurrence of neat morphological changes in the cellular shape.

A synchrotron-based infrared microspectroscopy study on the cellular response induced by gold nanoparticles combined with X-ray irradiations on F98 and U87-MG glioma cell lines

“…38 Changes in these peaks were previously related to the changes in the lipid structure and chain length due to oxidative stress and membrane peroxidation. [38][39][40][41] The separation in the PCA scores between −NP and +NP groups is less clear in the case of U87-MG cells (Fig. 4).…”

“…6 (bottom) show a clear increase in the CH 2 /CH 3 ratio distribution for F98 cells (both at T 0h and T 24h ), which might indicate evidences of oxidative stress. 39,40 There are also indications of oxidative stress in the U87-MG cells after NP-treatment (T 0h ), which seems to be reduced at T 24h . The induction of AuNP-related oxidative stress has been previously reported using the same type of NP via increased production of endogenous reactive oxygen species and depletion of intracellular antioxidants.…”

Biomolecular condensates undergo a generic shear-mediated liquid-to-solid transition

“…2a). By using AFM-IR, we acquired the maps of 3D morphology, IR absorption and stiffness of the FUS samples with nanoscale resolution 7,20,21 . We first acquired a morphology map of a single droplet (Fig.…”

Infrared Nanospectroscopy Reveals the Molecular Interaction Fingerprint of an Aggregation Inhibitor with Single Aβ42 Oligomers

“…1c). [24][25][26][27][28][29][30][31][32][33][34] AFM-IR is becoming widely applicable in biology since is capable of acquiring simultaneously morphological, nanomechanical and nanoscale-resolved chemical IR maps and absorption spectra from protein aggregates, liquid-liquid phase separated condensates, chromosomes, and single cells. 29,31,[34][35][36] As fundamental achievement, we have recently demonstrated that AFM-IR enables to acquire infrared absorption spectra for secondary structure determination from single protein molecules.…”