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.
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.
A dual optical tweezer has been built around an inverted microscope with high numerical aperture objective (N.A 1.4). The setup is versatile and can be used both as a single and a dual tweezer, and in the dual mode, enables us to optically trap two micron-sized latex beads within a few microns from each other in solution. Using this setup, we report measurements of the microrheological parameters of Pseudomonas°uorescens and Bacillus subtilis bacterial suspensions. We study the variation of viscoelastic moduli of these bacterial suspensions as a function of their cell count in solution. A comparison with inactive bacteria of corresponding cell count enables us to characterize the activity of the bacterial samples in terms of an average force that the bacteria exerts on the trapped bead. This work paves way for studies of interesting nonlinear rheological phenomena at small length scales.
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