4 March 2014 Optical properties of red blood cells: an optical tweezer based analysis
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A microscopic object finds an equilibrium orientation under a laser tweezer such that a maximum of its volume lies in the region of highest electric field. Furthermore, birefringent microscopic objects show no rotational diffusion after reorienting under a linearly polarized optical trap and also are seen to follow the plane of polarization when the latter is changed using a half wave plate. We observe that a healthy human Red Blood Cell (RBC) reproduces these observations in an optical tweezer, which confirms it to be birefringent. Polarization microscopy based measurements reveal that the birefringence is confined to the cell’s dimple region and the mean value of retardation for polarized green light (λ = 546nm) is 9 ± 1.5nm. We provide a simple geometrical model that attributes the birefringence to the nature of arrangement of the phospholipid molecules of the bilayer. This predicts the observed variation in the measured birefringence, from the dimple to the rim of the cell which we further show, can serve to demarcate the extent of the dimple region. This points to the value of birefringence measurements in revealing cell membrane contours. . We extend this technique to understand the birefringence of a chicken RBC, an oblate shaped cell, wherein the slow axis is identified to be coincident with the long axis of the cell. Further, we observe the birefringence to be confined to the edges of the cell. Experiments to probe the optomechanical response of the chicken RBC are in progress.
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B. V. Nagesh, B. V. Nagesh, Yogesha Lakkegowda, Yogesha Lakkegowda, R. Pratibha, R. Pratibha, P. Praveen, P. Praveen, Sarbari Bhattacharya, Sarbari Bhattacharya, Sharath Ananthamurthy, Sharath Ananthamurthy, "Optical properties of red blood cells: an optical tweezer based analysis", Proc. SPIE 8947, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII, 89471P (4 March 2014); doi: 10.1117/12.2039222; https://doi.org/10.1117/12.2039222

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