We report here on studies of reorientation of human red blood cells (RBCs) in an optical trap. We have measured the time required, tre, for the plane of the RBC entering the optical trap to undergo a 90-deg rotation to acquire an edge on orientation with respect to the beam direction. This has been studied as a function of laser power, P, at the trap center. The variation of tre with increasing P shows an initial sharp decrease followed by a much smaller rate of further decrease. We find that this experimentally measured variation is not in complete agreement with the variation predicted by a theoretical model where the RBC is treated as a perfectly rigid circular disk-like body. We argue that this deviation arises due to deformation of the RBC. We further reason that this feature is dominated by the elastic behavior of the RBC membrane. We compare the studies carried out on normal RBCs with RBCs where varying conditions of membrane stiffness are expected. We propose that the value of energy used for maximum deformation possible during a reorientation process is an indicator of the membrane elasticity of the system under study.
We report an enhancement in the corner frequency of an optically trapped non-magnetic microsphere in the plane perpendicular to the laser propagation direction on addition of ferrofluid to the suspension medium. We conjecture that a directed motion of the nanoparticles toward the trap in this plane is responsible for the augmentation. Changes in the corner frequency in the presence of external magnetic field gradients lend credence to this conjecture. Corner frequency augmentation is also observed when zinc oxide nanoparticles are used. Here, however, no further changes are seen in the presence of magnetic field gradients.
A normal human red blood cell (RBC) when trapped with a linearly polarized laser, reorients about the electric polarization direction and then remains rotationally bound to this direction. This behavior is expected for a birefringent object. We have measured the birefringence of distortion-free RBCs in an isotonic medium using a polarizing microscope. The birefringence is confined to the cell's dimple region and the slow axis is along a diameter. We report an average retardation of 3.5 ± 1.5 nm for linearly polarized green light (λ = 546 nm). We also estimate a retardation of 1.87 ± 0.09 nm from the optomechanical response of the RBC in an optical trap. We reason that the birefringence is a property of the cell membrane and propose a simple model attributing the origin of birefringence to the phospholipid molecules in the lipid bilayer and the variation to the membrane curvature. We observe that RBCs reconstituted in shape subsequent to crenation show diminished birefringence along with a sluggish optomechanical response in a trap. As the arrangement of phospholipid molecules in the cell membrane is disrupted on crenation, this lends credence to our conjecture on the origin of birefringence. Dependence of the birefringence on membrane contours is further illustrated through studies on chicken RBCs.
We have studied in detail the situation wherein two microbeads are trapped axially in a single-beam Gaussian intensity profile optical trap. We find that the corner frequency extracted from a power spectral density analysis of intensity fluctuations recorded on a quadrant photodetector (QPD) is dependent on the detection scheme. Using forward- and backscattering detection schemes with single and two laser wavelengths along with computer simulations, we conclude that fluctuations detected in backscattering bear true position information of the bead encountered first in the beam propagation direction. Forward scattering, on the other hand, carries position information of both beads with substantial contribution from the bead encountered first along the beam propagation direction. Mie scattering analysis further reveals that the interference term from the scattering of the two beads contributes significantly to the signal, precluding the ability to resolve the positions of the individual beads in forward scattering. In QPD-based detection schemes, detection through backscattering, thereby, is imperative to track the true displacements of axially trapped microbeads for possible studies on light-mediated interbead interactions.
Human red blood cells (RBCs) need to deform in order to pass through capillaries in human vasculature with diameter smaller than that of the RBC. An altered RBC cell membrane stiffness (CMS), thereby, is likely to have consequences on their flow rate. RBC CMS is known to be affected by several commonly encountered disease conditions. This study was carried out to investigate whether an increase in RBC CMS, to the extent seen in such commonly encountered medical conditions, affects the RBC flow rate through channels with diameters comparable to that of the RBC. To do this, we use RBCs extracted from a healthy individual with no known medical conditions and treated with various concentrations of Bovine Serum Albumin (BSA). We study their flow through polycarbonate membranes with pores of diameter 5μm and 8μm which are smaller than and comparable to the RBC diameter respectively. The studies are carried out at constant hematocrit and volumetric flow rate. We find that when the diameter of the capillary is smaller than that of the RBC, the flow rate of the RBCs is lowered as the concentration of BSA is increased while the reverse is true when the diameter is comparable to that of the RBC. We confirm that this is a consequence of altered CMS of the RBCs from their reorientation dynamics in an Optical Tweezer. We find that a treatment with 0.50mg/ml BSA mimics the situation for RBCs extracted from a healthy individual while concentrations higher than 0.50mg/ml elevate the RBC CMS across a range expected for individuals with a condition of hyperglycemia. Using a simple theoretical model of the RBC deformation process at the entry of a narrow channel, we extract the RBC membrane bending modulus from their flow rate.
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