Characterization Of Red Blood Cells Research Articles

Analysis of red blood cell deformability using parallel ladder electrodes in a microfluidic manipulation system

The mechanical properties of biological cells are known as effective label-free biomarkers in many cell physiological events. Several techniques have been proposed to manipulate, measure, and analyze cell mechanical properties, including micropipette aspiration, atomic force microscopy, optical tweezers, and magnetic tweezers. However, most of these methods have inherent problems in practice, including high equipment cost, complex operation, low accuracy, and low throughput. This paper presents a new method for the mechanical characterization of red blood cells (RBCs) on the basis of a dielectrophoresis (DEP) microfluidic system integrated with newly designed parallel ladder electrodes. The proposed microfluidic system not only can stretch RBCs in single-cell level exactly but also has the advantages of high efficiency and throughput. The DEP force generated by the ITO electrodes was theoretically studied, and a series of experiments were performed to stretch RBCs. The relationship between the DEP force and the strain of the RBCs was analyzed, which elucidated the elongation process of RBC under the domination of DEP force, as well as the maximum elongation ability of RBCs. These quantitative results are helpful for people to understand the characteristics of RBCs.

Bioinspired carbon quantum dots for sensitive fluorescent detection of vitamin B12 in cell system

Zwitterion-modification, as a bioinspired strategy, provides greatly promising platforms for biological detection and sensor applications. A green, low-cost and straight-forward method for synthesis of highly fluorescent biomimetic carbon quantum dots (BCQDs) has been developed via pyrolysis of cytidine diphosphate choline (CDPC) and ethylenediamine. The BCQDs with a strong emission at wavelength of 450 nm shows ultrasensitive sensing capability for vitamin B12 with high selectivity. Using the fluorometric assay, the detection limit (DL) for vitamin B12 was found to be as low as 81 nM. Meanwhile, the results of 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), hemolysis measuring and morphological characterization of Red blood cells (RBCs) confirms the excellent biocompatibility of BCQDs. The imaging experiments of human cervical cancer cells (HeLa) certify that BCQDs could be served as an effective fluorescent sensing probe for label-free sensitive and selective detection of vitamin B12 in biological samples on account of their low toxicity and good biocompatibility. The BCQDs, further, were successfully applied to probe vitamin B12 in living cells, which broaden its potential application in vivo system.

A Numerical Approach for Dielectrophoretic Characterization and Separation of Human Hematopoietic Cells

Rapid and reliable characterization of hematopoietic cells still remain the first step for precise medicine. Diagnosis of various diseases, ranging from infectious to cancer, relies on quantification of hematopoietic cells from blood. Therefore, there is an emerging need for label-free, lowcost, time-efficient, reproducible and quantitative characterization tools for the blood cells. Addressed herein is a numerical analysis for dielectrophoretic characterization of red blood cells, T-lymphocytes, B-lymphocytes and monocytes, which quantitatively incorporate with the membrane features of these cells to provide more insight into their dielectrophoretic responses.

Optical characterization of red blood cells from individuals with sickle cell trait and disease in Tanzania using quantitative phase imaging.

Sickle cell disease (SCD) is common across Sub-Saharan Africa. However, the investigation of SCD in this area has been significantly limited mainly due to the lack of research facilities and skilled personnel. Here, we present optical measurements of individual red blood cells from healthy individuals and individuals with SCD and sickle cell trait in Tanzania using the quantitative phase imaging technique. By employing a quantitative phase imaging unit, an existing microscope in a clinic is transformed into a powerful quantitative phase microscope providing measurements on the morphological, biochemical, and biomechanical properties of individual cells. The present approach will open up new opportunities for cost-effective investigation and diagnosis of several diseases in low resource environments.

Characterization of red blood cells with multiwavelength transmission spectroscopy.

Multiwavelength transmission (MWT) spectroscopy was applied to the investigation of the morphological parameters and composition of red blood cells (RBCs). The MWT spectra were quantitatively analyzed with a Mie theory based interpretation model modified to incorporate the effects of the nonsphericity and orientation of RBCs. The MWT spectra of the healthy and anemic samples were investigated for the RBC indices in open and blinded studies. When MWT performance was evaluated against a standard reference system, very good agreement between two methods, with R 2 > 0.85 for all indices studied, was demonstrated. The RBC morphological parameters were used to characterize three types of anemia and to draw an association between RBC morphology and anemia severity. The MWT spectra of RBCs infected with malaria parasite Plasmodium falciparum at different life cycle stages were analyzed for RBC morphological parameters. The changes in the RBC volume, surface area, aspect ratio, and hemoglobin composition were used to trace the morphological and compositional alterations in the infected RBCs occurring with parasites' development and to provide insights into parasite-host interactions. The MWT method was shown to be reliable for determination of the RBC morphological parameters and to be valuable for identification of the RBC pathologic changes and disease states.

High-throughput biophysical measurement of human red blood cells

This paper reports a microfluidic system for biophysical characterization of red blood cells (RBCs) at a speed of 100-150 cells s(-1). Electrical impedance measurement is made when single RBCs flow through a constriction channel that is marginally smaller than RBCs' diameters. The multiple parameters quantified as mechanical and electrical signatures of each RBC include transit time, impedance amplitude ratio, and impedance phase increase. Histograms, compiled from 84,073 adult RBCs (from 5 adult blood samples) and 82,253 neonatal RBCs (from 5 newborn blood samples), reveal different biophysical properties across samples and between the adult and neonatal RBC populations. In comparison with previously reported microfluidic devices for single RBC biophysical measurement, this system has a higher throughput, higher signal to noise ratio, and the capability of performing multi-parameter measurements.

Red blood cell proteomics

Since its discovery in the 17th century, the red blood cell, recognized in time as the critical cell component for survival, has been the focus of much attention. Its unique role in gas exchange (oxygen/CO 2 transport) and its distinct characteristics (absence of nucleus; biconcave cell shape) together with an – in essence – unlimited supply lead to extensive targeted biochemical, molecular and structural studies. A quick PubMed query with the word “erythrocyte” results in 198 013 scientific articles of which 162 are red blood cell proteomics studies, indicating that this new technique has been only recently applied to the red blood cell and related fields. Standard and comparative proteomics have been widely used to study different blood components. A growing body of proteomics literature has since developed, which deals with the characterization of red blood cells in health and disease. The possibility offered by proteomics to obtain a global snapshot of the whole red blood cell protein make-up, has provided unique insights to many fields including transfusion medicine, anaemia studies, intra-red blood cell parasite biology and translational research. While the contribution of proteomics is beyond doubt, a full red blood cell understanding will ultimately require, in addition to proteomics, lipidomics, glycomics, interactomics and study of post-translational modifications. In this review we will briefly discuss the methodology and limitations of proteomics, the contribution it made to the understanding of the erythrocyte and the advances in red blood cell-related fields brought about by comparative proteomics.

Manipulation and characterization of red blood cells with alternating current fields in microdevices.

The motion of a suspension of erythrocytes (red blood cells, RBCs) in response to a high-frequency alternating current (AC) field in a microfluidic device is examined with parallel and orthogonal electrode configurations to delineate the various fundamental driving forces. Cell repulsion from the platinum electrodes due to electrode polarization interacting with cell membrane polarizations is observed to be the strongest force acting on the particles in the first few seconds of field application. We exploit this strong repulsion to concentrate the bioparticles between the microelectrodes to amplify multiparticle aggregation phenomenon and dielectrophoretic (DEP) manipulation in a small and well-characterized region within the microfluidic device. Secondary motions include RBC pearl chain formation along field lines due to particle polarization followed by classical dielectrophoretic motion of the chains across field lines to regions of weaker field. These are driven by far weaker dipole-dipole and field-dipole interactions than the preliminary electrode repulsions. RBC chain length and total aggregated cells are presented for a variety of AC frequencies and are significantly amplified by the electrode repulsion. Motion of particles away from the polarized electrode is found to be species- and age-sensitive and can stand by itself as a promising identification and separation mechanism. In a 0.1 S/m isotonic phosphate buffer saline medium, we observe the largest cell mobilities at an optimal frequency of approximately 1 MHz, corresponding to the inverse diffusion time across the double layer of the cell and across the electrode's polarized layer. This suggests that the dielectric responses of both particles and electrodes in the low MHz frequency range are mostly determined by normal electromigration of ions from the bulk to their interfaces. Sensitivity to RBC age and species suggests that the surface proteins and membrane ion channels can affect the capacitance of the interface to accommodate the ions from the bulk. Such surface ion accumulation and polarization mechanisms are different from the classical dielectric theories. The resonant frequency of electrode polarization at around 1 MHz falls between positive and negative dielectrophoretic resonant frequency peaks - suggesting that the double-layer polarization mechanism is a distinct and potentially important bioparticle manipulation tool.

Venous and arterial hematologic profiles of very low birth weight infants. European Multicenter rhEPO Study Group.

In very low birth weight (VLBW) infants, diagnostic and therapeutic decisions depend on hematologic values. As few data are available, we studied the course during the first 6 weeks of life. Four prospective longitudinal cohort studies were retrospectively combined assessing hematologic profiles of 562 VLBW infants. For characterization of red blood cells and iron, infants receiving erythropoietin were excluded. For characterization of white blood cells and platelets, infants receiving antibiotics were excluded. The third (3rd)/median/97th percentiles on day 3 were as follows: hemoglobin: 11.0/15.6/19.8 g/dL; hematocrit: 35/47/60%; red blood cells: 3.2/4.2/5.3 x 10(12)/L; reticulocytes:. 6/7.1/27.8%; platelets: 58/203/430 x 10(9)/L; white blood cells: 3. 6/9.5/38.3 x 10(9)/L; neutrophils:.7/4.7/25.3 x 10(9)/L; ferritin: 27/140/504 ng/mL; iron:.8/7.5/26.7 micromol/L; transferrin saturation: 2.6/22.7/79.8%. Transferrin saturation was <24% in 51%, ferritin concentration <100 ng/mL in 32%, and platelets <150 x 10(9)/L in 29% of this population. The steady decrease of red cell parameters was mitigated by transfusions. Neutrophils decreased steadily, and were <1.75 x 10(9)/L in 35% at 6 weeks. Iron indices and platelet counts on day 3 and neutrophil counts at 2 to 6 weeks of age are lower than previously assumed in VLBW infants and lower than in larger prematures.

Separation and characterization of red blood cells with different membrane deformability using steric field-flow fractionation

Human red blood cells were treated in different ways to alter their membrane deformability, and the hydrodynamic behavior of these altered cells was studied using the steric field-flow fractionation (FFF) technique. The relationships between cell retention in the FFF channel, flow-rate of the carrier fluid and the applied field strength were studied for normal and glutaraldehyde-fixed human red cells, and separation conditions were optimized. The effect of flow-induced hydrodynamic lift forces on red cell retention in the steric FFF channel was studied, and the results suggest that the membrane deformability of the red cell is an important factor contributing to the lift force, besides other previously described effects due to density and flow velocity. Using steric FFF, a mixture of normal and glutaraldehyde-fixed human red cells was completely separated with a resolution twice that found in published d ata from gel permeation, another hydrodynamic separation technique. Partial loss of membrane deformability, induced by different degrees of glutaraldehyde-fixation, by diamide, or by a thermal treatment, has also been studied. Steric FFF is thus shown to have potential for rapid separation and differentiation of red cells with different density and membrane deformability, conditions known to be associated with, e.g., cell senescence and certain hematological diseases.

Separation and characterization of red blood cells from newborns and infants during the first trimenon of life using a dextran density gradient: mean cellular volume and mean cellular haemoglobin concentration.

The adaptation of cell physiological parameters of erythrocytes of the newborn to erythrocytes in later life was studied by cell separation in an isopycnic dextran density gradient. The cell characteristics during the first trimester of life were followed by repeated determinations. The mean cellular haemoglobin concentration of the lightest cells is remarkably less than that of the cells of median density. This is probably due to the ongoing synthesis of haemoglobin in reticulocytes, which are highly concentrated in the top fractions. It was shown that the young cells released from the bone marrow become progressively smaller during this period with a decrease of mean cellular volume (MCV) from 104 micrometer3 at birth to 85 micrometer3 at the end of the first trimester. It has been established that this process commences during the first week of life. No difference concerning the alterations of cell parameters, especially MCV, in premature and mature infants was found. It is therefore probable that these alterations are due to changes in the external milieu following birth.

Characterization of red blood cells strongly coated in vitro by C3 via the alternative pathway.

Characterization of red blood cells strongly coated in vitro by C3 via the alternative pathway.