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optical tweezers

Aaron Yevick, Daniel J Evans, David G Grier
The theory of photokinetic effects expresses the forces and torques exerted by a beam of light in terms of experimentally accessible amplitude and phase profiles. We use this formalism to develop an intuitive explanation for the performance of optical tweezers operating in the Rayleigh regime, including effects arising from the influence of light's angular momentum. First-order dipole contributions reveal how a focused beam can trap small objects, and what features limit the trap's stability. The first-order force separates naturally into a conservative intensity-gradient term that forms a trap and a non-conservative solenoidal term that drives the system out of thermodynamic equilibrium...
February 28, 2017: Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences
Ieva Ailte, Anne Berit Dyve Lingelem, Audun Sverre Kvalvaag, Simona Kavaliauskiene, Andreas Brech, Gerbrand Koster, Paul Gunnar Dommersnes, Jonas Bergan, Tore Skotland, Kirsten Sandvig
In this study, we have investigated how clathrin-dependent endocytosis is affected by exogenously added lysophospholipids (LPLs). Addition of LPLs with large head groups strongly inhibits transferrin (Tf) endocytosis in various cell lines, while LPLs with small head groups do not. Electron and total internal reflection fluorescence microscopy (EM and TIRF) reveal that treatment with lysophosphatidylinositol (LPI) with the fatty acyl group C18:0 leads to reduced numbers of invaginated clathrin-coated pits (CCPs) at the plasma membrane, fewer endocytic events per membrane area and increased lifetime of CCPs...
January 9, 2017: Traffic
M Hashemi Shabestari, A E C Meijering, W H Roos, G J L Wuite, E J G Peterman
Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that has found widespread application in biology. In this combined approach, the spatial (~nm) and temporal (~ms) resolution, as well as the force scale (~pN) accessible to optical tweezers is complemented with the power of fluorescence microscopy...
2017: Methods in Enzymology
J Wang, J T Barnett, M R Pollard, N M Kad
Fluorescence imaging is one of the cornerstone techniques for understanding how single molecules search for their targets on DNA. By tagging individual proteins, it is possible to track their position with high accuracy. However, to understand how proteins search for targets, it is necessary to elongate the DNA to avoid protein localization ambiguities. Such structures known as "DNA tightropes" are tremendously powerful for imaging target location; however, they lack information about how force and load affect protein behavior...
2017: Methods in Enzymology
K D Whitley, M J Comstock, Y R Chemla
We describe the design, construction, and application of an instrument combining dual-trap, high-resolution optical tweezers and a confocal microscope. This hybrid instrument allows nanomechanical manipulation and measurement simultaneously with single-molecule fluorescence detection. We present the general design principles that overcome the challenges of maximizing optical trap resolution while maintaining single-molecule fluorescence sensitivity, and provide details on the construction and alignment of the instrument...
2017: Methods in Enzymology
E Brost, J Brooks, J Piepenburg, S Chakraborty, T Das, A Green, Y Watanabe, S Hui
PURPOSE: Patients with BCR-ABL (Ph +ve) acute lymphoblastic leukemia are at very high risk of relapse and mortality. In line with the NIH mission to understand the physical and biological processes, we seek to report mechano-biological method to assessment and distinguish treated/untreated leukemia cells. METHODS: BCR-ABL leukemia cell populations and silica microspheres were trapped in a 100x magnification optical trapping system (λ=660 nm, 70 mW). Light refracted through the trapped sample was collected in the back focal plane by a quadrant detector to measure the positions of individual cells...
June 2016: Medical Physics
Yareni A Ayala, Bruno Pontes, Barbara Hissa, Ana Carolina M Monteiro, Marcos Farina, Vivaldo Moura-Neto, Nathan B Viana, H Moysés Nussenzveig
Mechanical properties of cells are known to be influenced by the actin cytoskeleton. In this article, the action of drugs that interact with the actin cortex is investigated by tether extraction and rheology experiments using optical tweezers. The influences of Blebbistatin, Cytochalasin D and Jasplakinolide on the cell mechanical properties are evaluated. The results, in contradiction to current views for Jasplakinolide, show that all three drugs and treatments destabilize the actin cytoskeleton, decreasing the cell membrane tension...
December 26, 2016: Experimental Cell Research
Zhi Zhu, Chaoyong James Yang
Heterogeneity among individual molecules and cells has posed significant challenges to traditional bulk assays, due to the assumption of average behavior, which would lose important biological information in heterogeneity and result in a misleading interpretation. Single molecule/cell analysis has become an important and emerging field in biological and biomedical research for insights into heterogeneity between large populations at high resolution. Compared with the ensemble bulk method, single molecule/cell analysis explores the information on time trajectories, conformational states, and interactions of individual molecules/cells, all key factors in the study of chemical and biological reaction pathways...
December 28, 2016: Accounts of Chemical Research
Karl Otto Greulich
The use of laser microbeams and optical tweezers in a wide field of biological applications from genomic to immunology is discussed. Microperforation is used to introduce a well-defined amount of molecules into cells for genetic engineering and optical imaging. The microwelding of two cells induced by a laser microbeam combines their genetic outfit. Microdissection allows specific regions of genomes to be isolated from a whole set of chromosomes. Handling the cells with optical tweezers supports investigation on the attack of immune systems against diseased or cancerous cells...
December 23, 2016: Reports on Progress in Physics
Zhihai Liu, Lu Liu, Zongda Zhu, Yu Zhang, Yong Wei, Xiaonan Zhang, Enming Zhao, Yaxun Zhang, Jun Yang, Libo Yuan
We propose and demonstrate a whispering gallery mode (WGM) resonance-based temperature sensor, where the microresonator is made of a DCM (2-[2-[4-(dimethylamino)phenyl] ethenyl]-6-methyl-4H-pyran-4-ylidene)-doped oil droplet (a liquid material) immersed in the water solution. The oil droplet is trapped, controlled, and located by a dual-fiber optical tweezers, which prevents the deformation of the liquid droplet. We excite the fluorescence and lasing in the oil droplet and measure the shifts of the resonance wavelength at different temperatures...
October 15, 2016: Optics Letters
Zhensheng Zhong, Lixia Yang, Haiping Zhang, Jiahao Shi, J Jeya Vandana, Do Thuy Uyen Ha Lam, René C L Olsthoorn, Lanyuan Lu, Gang Chen
Minus-one ribosomal frameshifting is a translational recoding mechanism widely utilized by many RNA viruses to generate accurate ratios of structural and catalytic proteins. An RNA pseudoknot structure located in the overlapping region of the gag and pro genes of Simian Retrovirus type 1 (SRV-1) stimulates frameshifting. However, the experimental characterization of SRV-1 pseudoknot (un)folding dynamics and the effect of the base triple formation is lacking. Here, we report the results of our single-molecule nanomanipulation using optical tweezers and theoretical simulation by steered molecular dynamics...
December 21, 2016: Scientific Reports
Zhuo Long Zhou, Jing Ma, Ming-Hui Tong, Barbara Pui Chan, Alice Sze Tsai Wong, Alfonso Hing Wan Ngan
The adhesion and traction behavior of leukemia cells in their microenvironment is directly linked to their migration, which is a prime issue affecting the release of cancer cells from the bone marrow and hence metastasis. In assessing the effectiveness of phorbol 12-myristate 13-acetate (PMA) treatment, the conventional batch-cell transwell-migration assay may not indicate the intrinsic effect of the treatment on migration, since the treatment may also affect other cellular behavior, such as proliferation or death...
2016: International Journal of Nanomedicine
Jochen Fick
Optical trapping of micron-size dielectric particles in a dual-fiber tip configuration is presented. Trap oscillation and suspension flow experiments are performed to investigate the linearity of the optical forces. These measurements are completed by standard methods such as Boltzmann statistics or power spectra evaluation. Strong trapping efficiencies of 0.25 and 1.8  pN·μm<sup>-1</sup> have been found in the axial and transverse directions, respectively. The values obtained by the different approaches are in good agreement...
December 15, 2016: Optics Letters
A Bérut, A Imparato, A Petrosyan, S Ciliberto
We analytically study the role of nonconservative forces, namely viscous couplings, on the statistical properties of the energy flux between two Brownian particles kept at different temperatures. From the dynamical model describing the system, we identify an energy flow that satisfies a fluctuation theorem both in the stationary and in transient states. In particular, for the specific case of a linear nonconservative interaction, we derive an exact fluctuation theorem that holds for any measurement time in the transient regime, and which involves the energy flux alone...
November 2016: Physical Review. E
Roman D Bulushev, Sanjin Marion, Ekaterina Petrova, Sebastian J Davis, Sebastian J Maerkl, Aleksandra Radenovic
Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding sites on λ-DNA (48.5 kbp). Measured localization of binding sites showed a shift from the expected positions on the DNA that we explained using both analytical fitting and a stochastic model. From the measured force versus stage curves we extracted the nonequilibrium work done during the translocation of a DNA-protein complex and used it to obtain an estimate of the effective charge of the complex...
December 14, 2016: Nano Letters
Julian Hering, Erik H Waller, Georg Von Freymann
Controlling the point-spread-function in three-dimensional laser lithography is crucial for fabricating structures with highest definition and resolution. In contrast to microscopy, aberrations have to be physically corrected prior to writing, to create well defined doughnut modes, bottlebeams or multi foci modes. We report on a modified Gerchberg-Saxton algorithm for spatial-light-modulator based automated aberration compensation to optimize arbitrary laser-modes in a high numerical aperture system. Using circularly polarized light for the measurement and first-guess initial conditions for amplitude and phase of the pupil function our scalar approach outperforms recent algorithms with vectorial corrections...
December 12, 2016: Optics Express
Robin Diekmann, Deanna L Wolfson, Christoph Spahn, Mike Heilemann, Mark Schüttpelz, Thomas Huser
Imaging non-adherent cells by super-resolution far-field fluorescence microscopy is currently not possible because of their rapid movement while in suspension. Holographic optical tweezers (HOTs) enable the ability to freely control the number and position of optical traps, thus facilitating the unrestricted manipulation of cells in a volume around the focal plane. Here we show that immobilizing non-adherent cells by optical tweezers is sufficient to achieve optical resolution well below the diffraction limit using localization microscopy...
December 13, 2016: Nature Communications
An-Ping Yu, Gang Chen, Zhi-Hai Zhang, Zhong-Quan Wen, Lu-Ru Dai, Kun Zhang, Sen-Lin Jiang, Zhi-Xiang Wu, Yu-Yan Li, Chang-Tao Wang, Xian-Gang Luo
The generation of a sub-diffraction longitudinally polarized spot is of great interest in various applications, such as optical tweezers, super-resolution microscopy, high-resolution Raman spectroscopy, and high-density optical data storage. Many theoretical investigations have been conducted into the tight focusing of a longitudinally polarized spot with high-numerical-aperture aplanatic lenses in combination with optical filters. Optical super-oscillation provides a new approach to focusing light beyond the diffraction limit...
December 12, 2016: Scientific Reports
Vishnu Chandra, Zain Hannan, Huizhong Xu, Maumita Mandal
Guanine-responsive riboswitches undergo ligand-dependent structural rearrangements to control gene expression by transcription termination. While the molecular basis for ligand recognition is well established, the associated structural rearrangements and the kinetics involved in the formation of the aptamer domain are less well understood. Using high-resolution optical tweezers, we followed the folding trajectories of a single molecule of the xpt-pbuX guanine aptamer from Bacillus subtilis. We report a rapid six-state conformational rearrangement, in which three of the states are guanine dependent, during the transition from the linear to the native receptor conformation...
December 12, 2016: Nature Chemical Biology
Rui-Qiong Luo, Fang Wei, Shu-Shi Huang, Yue-Ming Jiang, Shan-Lei Zhang, Wen-Qing Mo, Hong Liu, Xi Rong
The examination of insulin (Ins) exocytosis at the single-cell level by conventional methods, such as electrophysiological approaches, total internal reflection imaging, and two-photon imaging technology, often requires an invasive microelectrode puncture or label. In this study, high concentrations of glucose and potassium chloride were used to stimulate β cell Ins exocytosis, while low concentrations of glucose and calcium channel blockers served as the blank and negative control, respectively. Laser tweezers Raman spectroscopy (LTRS) was used to capture the possible Raman scattering signal from a local zone outside of the cell edge...
December 9, 2016: Applied Spectroscopy
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