Characterizing the rotation of non symmetric objects in an optical tweezer

We present an optical tweezer based study of the rotation of microscopic objects with shape asymmetry. Thermal fluctuations and rotations are simultaneously monitored through laser back scattering. The rotation causes a modulation in intensity of the back scattered light incident on a quadrant photo detector. The resulting power spectrum is a modified Lorentzian with additional peaks located at the fundamental rotational frequency of the object and at the integer harmonics. The manifestation of these peaks reveals that the rotations are periodic but with varying angular velocity. We model our experimental results to illustrate the hydrodynamic interplay between the rotor and the surrounding medium that results in the time dependence of the angular speed of the former. Further, we demonstrate the use of video microscopy for characterization of low reflectivity rotors, such as biological cells. We propose through these studies that an analysis of these rotations can provide insights into the role of hydrodynamics at micron levels

An optical tweezer-based study of antimicrobial activity of silver nanoparticles

Understanding and characterizing microbial activity reduction in the presence of antimicrobial agents can help in the design and manufacture of antimicrobial drugs. We demonstrate the use of an optical tweezer setup in recording the changes in bacterial activity with time, induced by the presence of foreign bodies in a bacterial suspension. This is achieved by monitoring the fluctuations of an optically trapped polystyrene bead immersed in it. Examining the changes in the fluctuation pattern of the bead with time provides an accurate characterization of the reduction in the microbial activity. Here, we report on the effect of addition of silver nanoparticles on bacterial cultures of Pseudomonas aeroginosa, Escherichia coli and Bacillus subtilis. We observe a decrease in the bacterial activity with time for the investigated bacterial samples. This method in our opinion, enables one to track changes in bacterial activity levels as a function of time of contact with the antibacterial agent with greater efficacy than traditional cell counting methods. © Indian Academy of Sciences

A Dual Optical Tweezer for Microrheology of Bacterial Suspensions

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 soln. Using this setup, we report measurements of the microrheol. parameters of Pseudomonas fluorescens and Bacillus subtilis bacterial suspensions. We study the variation of viscoelastic moduli of these bacterial suspensions as a function of their cell count in soln. A comparison with inactive bacteria of corresponding cell count enables us to characterize the activity of the bacterial samples in terms of an av. force that the bacteria exerts on the trapped bead. This work paves way for studies of interesting nonlinear rheol. phenomena at small length scales

A Helix Replacement Mechanism Directs Metavinculin Functions

Cells require distinct adhesion complexes to form contacts with their neighbors or the extracellular matrix, and vinculin links these complexes to the actin cytoskeleton. Metavinculin, an isoform of vinculin that harbors a unique 68-residue insert in its tail domain, has distinct actin bundling and oligomerization properties and plays essential roles in muscle development and homeostasis. Moreover, patients with sporadic or familial mutations in the metavinculin-specific insert invariably develop fatal cardiomyopathies. Here we report the high resolution crystal structure of the metavinculin tail domain, as well as the crystal structures of full-length human native metavinculin (1,134 residues) and of the full-length cardiomyopathy-associated ΔLeu954 metavinculin deletion mutant. These structures reveal that an α-helix (H1′) and extended coil of the metavinculin insert replace α-helix H1 and its preceding extended coil found in the N-terminal region of the vinculin tail domain to form a new five-helix bundle tail domain. Further, biochemical analyses demonstrate that this helix replacement directs the distinct actin bundling and oligomerization properties of metavinculin. Finally, the cardiomyopathy associated ΔLeu954 and Arg975Trp metavinculin mutants reside on the replaced extended coil and the H1′ α-helix, respectively. Thus, a helix replacement mechanism directs metavinculin's unique functions

Optical tweezer for micro and nano scale rheology of biomaterials

Since the first proposal and demonstration of an optical trap to hold micron sized particles by Ashkin in 1986, optical tweezer-based techniques for control and manipulation of particles has seen tremendous progress and enabled the exploration of the structure and dynamics of materials at the nano length scale. In biophysical applications, these techniques have enabled a range of activities from measurements of very small forces (at sub-pN levels) at operation within cells thus assisting in the elucidation of intracellular transport mechanisms, to obtaining elastic parameters of biomolecules. The techniques have helped in gaining insights on the viscoelastic properties at micro and nano length scales of a range of soft materials such as colloids, polymer melts, bio membranes etc. The paper describes some of the studies carried out by us to understand polymer structure and elasticity of these materials in solution. The domains of "passive" and "active" rheology are explained with examples of studies undertaken to address these domains. The work using video microscopy to study silk fibroin solutions is used to exemplify the domain of passive rheology. We present recent work on bacterial solutions wherein live bacteria influence the optical trap region and thereby help us study the local viscoelasticity of such a medium. © 2010 IACS

Dynamics of a pair of trapped beads in a single beamoptical tweezer

We attempt to understand the interaction between a pair of trapped beads in a single beam tweezer by studying the difference in the corner frequencies and by estimating the correlation coefficient between them. © 2013 The Optical Society (OSA)

Cross-Talk of Phosphorylation and Prolyl Isomerization of the C-terminal Domain of RNA Polymerase II

Post-translational modifications of the heptad repeat sequences in the C-terminal domain (CTD) of RNA polymerase II (Pol II) are well recognized for their roles in coordinating transcription with other nuclear processes that impinge upon transcription by the Pol II machinery; and this is primarily achieved through CTD interactions with the various nuclear factors. The identification of novel modifications on new regulatory sites of the CTD suggests that, instead of an independent action for all modifications on CTD, a combinatorial effect is in operation. In this review we focus on two well-characterized modifications of the CTD, namely serine phosphorylation and prolyl isomerization, and discuss the complex interplay between the enzymes modifying their respective regulatory sites. We summarize the current understanding of how the prolyl isomerization state of the CTD dictates the specificity of writers (CTD kinases), erasers (CTD phosphatases) and readers (CTD binding proteins) and how that correlates to transcription status. Subtle changes in prolyl isomerization states cannot be detected at the primary sequence level, we describe the methods that have been utilized to investigate this mode of regulation. Finally, a general model of how prolyl isomerization regulates the phosphorylation state of CTD, and therefore transcription-coupled processes, is proposed

Correlation between microstructure and microrheological parameters of various silk fibres

The changes in microcrystalline parameters of raw wild varieties of silk fibres, like tasar, muga and eri, have been studied using wide angle X-ray scattering technique and a line profile analysis. A method involving an exponential distribution has been used to compute the microstructural parameters for the crystallite. In addition, a home-built open microscope set-up is also used for determining the microrheological parameters for all the three silk varieties in solution form. A comparative study reveals interesting correlations in the relative strengths of the varieties of silk fibres in both crystalline form and in solution. Further, the findings also reveal that muga is stiffer than the other non-mulberry silk varieties and this is observed in both the forms