Probing orientation and rotation of red blood cells in optical tweezers by digital holographic microscopy

Abstract

Interaction of red blood cells (RBC) with optical tweezers has been found to differ under varied physiological and pathological conditions as compared to its normal conditions. Earlier, we reported difference in rotation of trapped RBC in hypertonic conditions for detection of malaria infection. Disk-like RBC when trapped in optical tweezers get oriented in the vertical plane to maximize interaction with trapping beam. However, classical bright field, phase contrast or epifluorescence microscopy cannot confirm its orientation, thus leading to ambiguous conclusions such as folding of RBC during trapping by some researchers. Now, with use of digital holographic microscopy (DHM), we achieved high axial sensitivity that confirmed orientation of trapped red blood cell. Further, DHM enabled quantitative phase imaging of RBC under hypertonic condition. Dynamic changes of rotating RBC under optical tweezers at different trapping laser power were evaluated by the use of DHM. The deviation from linear dependence of rotation speed of RBC on laser power, was attributed towards deformation of RBC shape due to higher laser power (or speed).