Shruthi S. Iyengar

Bio: Shruthi S. Iyengar is an academic researcher from Bangalore University. The author has contributed to research in topics: Optical tweezers & Birefringence. The author has an hindex of 5, co-authored 13 publications receiving 45 citations.

Orientational dynamics of human red blood cells in an optical trap.

Abstract: We report here on studies of reorientation of human red blood cells (RBCs) in an optical trap. We have measured the time required, t re , 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 t re 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.

Birefringence of a normal human red blood cell and related optomechanics in an optical trap.

Abstract: 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.

Estimation of membrane bending modulus of stiffness tuned human red blood cells from micropore filtration studies

Abstract: 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.

Two particle tracking and detection in a single Gaussian beam optical trap.

Abstract: 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.

Conical Fresnel zone lens for optical trapping

Abstract: The phase of a negative axicon is combined with that of a Fresnel zone lens (FZL) to obtain an element labelled as conical FZL, which can generate a focused ring pattern at the focal plane of the FZL. The phase integration is achieved by modifying the location and width of zones of FZL in accordance with the phase variation of the negative axicon. The element was designed for a high power laser with a wavelength of 1064 nm, focal length and diameter of conical FZL of 30 mm and 8 mm respectively and for a ring diameter of 50 μm. The element was fabricated using photolithography. The pattern was transferred from the resist layer to the borosilicate glass plates by dry etching to achieve an etch depth of 1064 nm. The etch depth measured using confocal microscope was 1034 nm at the central part and 930 nm for the outermost part of the device with a maximum error of 12.5% at the outermost part and 3% at the central part. The element was used in an optical trapping experiment. The ring pattern generated by the conical FZL was reimaged into the trapping plane using a tightly focusing microscopic objective. Polystyrene beads with diameters of 3 μm were suspended in deionized distilled water at the trapping plane. The element was found to trap multiple particles in to the same trap.

Optical alignment and spinning of laser-trapped microscopic particles

Abstract: Light-induced rotation of absorbing microscopic particles by transfer of angular momentum from light to the material raises the possibility of optically driven micromachines. The phenomenon has been observed using elliptically polarized laser beams or beams with helical phase structure. But it is difficult to develop high power in such experiments because of overheating and unwanted axial forces, limiting the achievable rotation rates to a few hertz. This problem can in principle be overcome by using transparent particles, transferring angular momentum by a mechanism first observed by Beth in 1936, when he reported a tiny torque developed in a quartz waveplate due to the change in polarization of transmitted light. Here we show that an optical torque can be induced on microscopic birefringent particles of calcite held by optical tweezers. Depending on the polarization of the incident beam, the particles either become aligned with the plane of polarization (and thus can be rotated through specified angles) or spin with constant rotation frequency. Because these microscopic particles are transparent, they can be held in three-dimensional optical traps at very high power without heating. We have observed rotation rates in excess of 350 Hz.

Nondestructive Identification and Accurate Isolation of Single Cells through a Chip with Raman Optical Tweezers.

Abstract: Raman optical tweezers (ROT) as a label-free technique plays an important role in single-cell study such as heterogeneity of tumor and microbial cells. Herein we designed a chip utilizing ROT to isolate a specific single cell. The chip was made from a polydimethylsiloxane (PDMS) slab and formed into a gourd-shaped reservoir with a connected channel on a cover glass. On the chip an individual cell could be isolated from a cell crowd and then extracted with ∼0.5 μL of phosphate-buffered saline (PBS) via pipet immediately after Raman spectral measurements of the same cell. As verification, we separated four different type of cells including BGC823 gastric cancer cells, erythrocytes, lymphocytes, and E. coli cells and quantifiably characterized the heterogeneity of the cancer cells, leukocyte subtype, and erythrocyte status, respectively. The average time of identifying and isolating a specific cell was 3 min. Cell morphology comparison and viability tests showed that the successful rate of single-cell isolation was about 90%. Thus, we believe our platform could further couple other single-cell techniques such as single-cell sequencing and become a multiperspective analytical approach at the level of a single cell.

Raman spectroscopic investigations on optical trap induced deoxygenation of red blood cells

Abstract: Raman spectroscopic investigations on the oxygenation status of optically trapped red blood cells show that the cellular site in the trap beam is more deoxygenated compared to the rest of the cell, and the level of deoxygenation increases with an increase in the trap beam power These observations and the changes in the Raman spectrum of hemoglobin solution as a function of the trapping beam power suggest that observed deoxygenation may be due to photodissociation of oxygen from hemoglobin at increased trapping power

Experimental demonstration of square Fresnel zone plate with chiral side lobes.

Abstract: In this study, we introduce what we believe is a novel holographic optical element called a chiral square Fresnel zone plate (CSFZP) The chirality is imposed on a square Fresnel zone plate (SFZP) using a nonclassical technique by rotating the half-period zones relative to one another The rotation of the half-period zones, in turn, twists the side lobes of the diffraction pattern without altering the focusing properties inherent to a SFZP As a consequence, the beam profile is hybrid, consisting of a strong central Gaussian focal spot with gradient force similar to that generated by a lens and twisted side lobes with orbital angular momentum The optical fields at the focal plane were calculated and found to possess a whirlpool-phase profile and a twisted intensity profile Analysis of the field variation along the direction of propagation revealed a spiraling phase and amplitude distribution Poynting vector plot of the fields revealed the presence of angular momentum in the regions of chiral side lobes The phase of the CSFZPs were displayed on a phase-only reflective spatial light modulator and illuminated using a laser The intensity patterns recorded in the experiment match the calculated ones, with a strong central focal spot and twisted side lobes The beam pattern was implemented in an optical trapping experiment and was found to possess particle trapping capabilities