Zhou, Zhaokun, Miller, Helen, Wollman, Adam orcid.org/0000-0002-5501-8131 et al. (1 more author) (2015) Developing a new biophysical tool to combine magneto-optical tweezers with super-resolution fluorescence microscopy. Photonics. pp. 758-772. ISSN 2304-6732
Abstract
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | This content is made available by the publisher under a Creative Commons Attribution Licence. This means that a user may copy, distribute and display the resource providing that they give credit. Users must adhere to the terms of the licence. |
Keywords: | magnetic tweezers,optical tweezers,super-resolution microscopy |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) The University of York > Faculty of Sciences (York) > Physics (York) |
Depositing User: | Pure (York) |
Date Deposited: | 21 Jul 2015 14:18 |
Last Modified: | 04 Dec 2024 00:11 |
Published Version: | https://doi.org/10.3390/photonics2030758 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.3390/photonics2030758 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:87399 |
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