Role of Arbitrary Intensity Profile Laser Beam in Trapping of RBC for Phase-imaging

Abstract

Red blood cells (RBCs) are customarily adhered to a bio-functionalised substrate to make them stationary in interferometric phase-imaging modalities. This can make them susceptible to receive alterations in innate morphology due to their own weight. Optical tweezers (OTs) often driven by Gaussian profile of a laser beam is an alternative modality to overcome contact-induced perturbation but at the same time a steeply focused laser beam might cause photo-damage. In order to address both the photo-damage and substrate adherence induced perturbations, we were motivated to stabilize the RBC in OTs by utilizing a laser beam of ‘arbitrary intensity profile’ generated by a source having cavity imperfections per se. Thus the immobilized RBC was investigated for phase-imaging with sinusoidal interferograms generated by a compact and robust Michelson interferometer which was designed from a cubic beam splitter having one surface coated with reflective material and another adjacent coplanar surface aligned against a mirror. Reflected interferograms from bilayers membrane of a trapped RBC were recorded and analyzed. Our phase-imaging set-up is limited to work in reflection configuration only because of the availability of an upright microscope. Due to RBC’s membrane being poorly reflective for visible wavelengths, quantitative information in the signal is weak and therefore, the quality of experimental results is limited in comparison to results obtained in transmission mode by various holographic techniques reported elsewhere.