Significance: We introduce a model for better calibration of the trapping force using an equal but oppositely directed drag force acting on a trapped red blood cell (RBC). We demonstrate this approach by studying RBCs’ elastic properties from deidentified sickle cell anemia (SCA) and sickle cell trait (SCT) blood samples.
Aim: A laser trapping (LT) force was formulated and analytically calculated in a cylindrical model. Using this trapping force relative percent difference, the maximum (longitudinal) and minimum (transverse) radius rate and stiffness were used to study the elasticity.
Approach: The elastic property of SCA and SCT RBCs was analyzed using LT technique with computer controlled piezo-driven stage, in order to trap and stretch the RBCs.
Results: For all parameters, the results show that the SCT RBC samples have higher elastic property than the SCA RBCs. The higher rigidity in the SCA cell may be due to the lipid composition of the membrane, which was affected by the cholesterol concentration.
Conclusions: By developing a theoretical model for different trapping forces, we have also studied the elasticity of RBCs in SCT (with hemoglobin type HbAS) and in SCA (with hemoglobin type HbSS). The results for the quantities describing the elasticity of the cells consistently showed that the RBCs in the SCT display lower rigidity and higher deformability than the RBCs with SCA.
An approach to an established technique that is potentially applicable for a more comprehensive understanding of the electrical properties of red blood cells (RBCs) is presented. Using a high-intensity gradient laser trap, RBCs can be singly trapped and consequentially ionized. The subsequent dynamics of the ionized cell allows one to calculate the charge developed and the ionization energy (IE) through a Newtonian-based analysis. RBCs with two different hemoglobin (Hb) types were ionized. The first sample was identified as carrying Hb HbAA (normal Hb) and the second one was identified as carrying HbAC (HbC trait). By analyzing the charge developed on each cell and several other related factors, we were able to discern a difference between the main Hb types contained within the individual RBC, independent of cell size. A relationship between the charge developed and the IE of the cell was also established based on the electrical properties of RBCs. Thus, we present this laser trapping technique as a study of the electrical properties of RBCs and as possible biomedical tool to be used for the differentiation of Hb types.
Rance Solomon, James Cooper, Gabriel Welker, Elaura Aguilar, Brooke Flanagan, Chelsey Pennycuff, David Scott, Anthony Farone, Mary Farone, Daniel Erenso, Robert Mushi, Maria del Pilar Aguinaga
Genetic mutation of the β-globin gene or inheritance of this mutated gene changes the chemical composition of the oxygen-carrying hemoglobin molecule that could lead to either the heterozygote genotype, resulting in sickle cell trait (SCT), or the homozygote genotype, resulting in sickle cell anemia (SCA). These mutations could affect the reversible elastic deformations of the red blood cells (RBCs) which are vital for biological functions. We have investigated this effect by studying the differences in the deformability of RBCs from blood samples of an individual with SCT and an untreated patient with SCA along with hemoglobin quantitation of each blood sample. Infrared 1064 nm laser trap force along with drag shear force are used to induce deformation in the RBCs. Ultra2-High Performance Liquid Chromatography (UHPLC) is used for the hemoglobin quantitation.
A laser tweezer (LT) along with advanced imaging techniques has been widely applied to manipulate and study living as well as nonliving microscopic objects. In this study we present yet another novel application of LTs for a precise measurement of the viscosities of fluids in a micro-volume flow. We have demonstrated this novel application by measuring the viscosity of a fetal bovine serum (FBS) using a LT constructed from a single intensity gradient laser trap. By calibrating the LT using dielectric silica micro-beads in a fluid with a known viscosity, specifically water, and by suspending same size of silica beads in the FBS and trapping with the same trap, we have determined the viscosity of the FBS at different temperatures. We have used the relationship between the trapping and Stoke’s drag force for a constant drag speed to determine the viscosity. We have also analyzed the viscosities determined in comparison with corresponding viscosities measured using an Ostwald viscometer.
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