Image- and Fluorescence-Based Test Shows Oxidant-Dependent Damages in Red Blood Cells and Enables Screening of Potential Protective Molecules

An increase of oxygen saturation within blood bags and metabolic dysregulation occur during storage red cells (RBCs). It leads to the gradual exhaustion RBC antioxidant protective system and, consequently, a deleterious state oxidative stress that plays major role in apparition so-called lesions. The present study describes use test (called TSOX) based on fluorescence label-free morphology readouts simply quickly evaluate oxidant properties various compounds controlled conditions. Here, TSOX was applied RBCs treated with four antioxidants (ascorbic acid, uric trolox resveratrol) three oxidants (AAPH, diamide H2O2) at different concentrations. Two complementary were chosen: first, where ROS generation quantified using DCFH-DA fluorescent probe, second, digital holographic microscopy measures alterations. All produced an fluorescence, whereas H2O2 did not visibly impact morphology. Significant protection observed out added molecules. Of note, resveratrol induced diamond-shape “Tirocytes”. assay design selected be flexible, as well compatible high-throughput screening. In future experiments, will serve screen chemical libraries probe molecules could additive solution for storage.