Developing a New Biophysical Tool to Combine Magneto-Optical Tweezers with Super-Resolution Fluorescence Microscopy

Zhaokun Zhou, Helen Miller, Adam J.M. Wollman and Mark C. Leake

We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and manipulation. Previously we have developed a super-resolution imaging module which, in conjunction with advanced imaging techniques such as Blinking assisted Localisation Microscopy (BaLM), achieves localisation precision of single fluorescent dye molecules bound to DNA of ~30 nm along the contour of the molecule; our work here describes developments in producing a system which combines tweezing and super-resolution fluorescence imaging. The instrument also features an acousto-optic deflector that temporally divides the laser beam to form multiple traps for high throughput statistics collection. Our motivation for developing the new tool is to enable direct observation of detailed molecular topological transformation and protein binding event localisation in a stretching/twisting mechanical assay that previously could hitherto only be deduced indirectly from the end-to-end length variation of DNA. Our approach is simple and robust enough for reproduction in the lab without the requirement of precise hardware engineering, yet is capable of unveiling the elastic and dynamic properties of filamentous molecules that have been hidden using traditional tools.

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Optical Nanofiber Integrated into Optical Tweezers for In Situ Fiber Probing and Optical Binding Studies

Ivan Gusachenko, Viet Giang Truong, Mary C. Frawley and Síle Nic Chormaic

Precise control of particle positioning is desirable in many optical propulsion and sorting applications. Here, we develop an integrated platform for particle manipulation consisting of a combined optical nanofiber and optical tweezers system. We show that consistent and reversible transmission modulations arise when individual silica microspheres are introduced to the nanofiber surface using the optical tweezers. The observed transmission changes depend on both particle and fiber diameter and can be used as a reference point for in situ nanofiber or particle size measurement. Thence, we combine scanning electron microscope (SEM) size measurements with nanofiber transmission data to provide calibration for particle-based fiber assessment. This integrated optical platform provides a method for selective evanescent field manipulation of micron-sized particles and facilitates studies of optical binding and light-particle interaction dynamics.

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Optical Fiber Tweezers Fabricated by Guided Wave Photo-Polymerization

Rita S. Rodrigues Ribeiro, Raquel Queirós, Olivier Soppera, Ariel Guerreiro and Pedro A. S. Jorge

In this work the use of guided wave photo-polymerization for the fabrication of novel polymeric micro tips for optical trapping is demonstrated. It is shown that the selective excitation of linear polarized modes, during the fabrication process, has a direct impact on the shape of the resulting micro structures. Tips are fabricated with modes LP02 and LP21 and their shapes and output intensity distribution are compared. The application of the micro structures as optical tweezers is demonstrated with the manipulation of yeast cells.

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A Model for the Force Exerted on a Primary Cilium by an Optical Trap and the Resulting Deformation

Ian Lofgren and Andrew Resnick

Cilia are slender flexible structures extending from the cell body; genetically similar to flagella. Although their existence has been long known, the mechanical and functional properties of non-motile (“primary”) cilia are largely unknown. Optical traps are a non-contact method of applying a localized force to microscopic objects and an ideal tool for the study of ciliary mechanics. We present a method to measure the mechanical properties of a cilium using an analytic model of a flexible, anchored cylinder held within an optical trap. The force density is found using the discrete-dipole approximation. Utilizing Euler-Bernoulli beam theory, we then integrate this force density and numerically obtain the equilibrium deformation of the cilium in response to an optical trap. The presented results demonstrate that optical trapping can provide a great deal of information and insight about the properties and functions of the primary cilium.

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Chirality in Optical Trapping and Optical Binding

David S. Bradshaw , Kayn A. Forbes , Jamie M. Leeder and David L. Andrews

Optical trapping is a well-established technique that is increasingly used on biological substances and nanostructures. Chirality, the property of objects that differ from their mirror image, is also of significance in such fields, and a subject of much current interest. This review offers insight into the intertwining of these topics with a focus on the latest theory. Optical trapping of nanoscale objects involves forward Rayleigh scattering of light involving transition dipole moments; usually these dipoles are assumed to be electric although, in chiral studies, magnetic dipoles must also be considered. It is shown that a system combining optical trapping and chirality could be used to separate enantiomers. Attention is also given to optical binding, which involves light induced interactions between trapped particles. Interesting effects also arise when binding is combined with chirality.

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