Abstract
The application of optical traps has come to the fore in the last three decades. They provide a powerful, sterile and noninvasive tool for the manipulation of cells, single biological macromolecules, colloidal microparticles and nanoparticles. An optically trapped microsphere may act as a force transducer that is used to measure forces in the piconewton regime. By setting up a well-calibrated single-beam optical trap within a fluorescence microscope system, one can measure forces and collect fluorescence signals upon biological systems simultaneously. In this protocol, we aim to provide a clear exposition of the methodology of assembling and operating a single-beam gradient force trap (optical tweezers) on an inverted fluorescence microscope. A step-by-step guide is given for alignment and operation, with discussion of common pitfalls.
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Acknowledgements
This work is supported by the UK Engineering and Physical Sciences Research Council. We acknowledge several useful discussions with Daniel Burnham.
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Supplementary information
Supplementary Figure 1
Sample Chamber (PDF 17 kb)
Supplementary Note 1
Dual beam optical tweezers system (PDF 90 kb)
Supplementary Note 2
Sample preparation (PDF 78 kb)
Supplementary Note 3
Q values (PDF 259 kb)
Supplementary Note 4
Trap stiffness (PDF 373 kb)
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Lee, W., Reece, P., Marchington, R. et al. Construction and calibration of an optical trap on a fluorescence optical microscope. Nat Protoc 2, 3226–3238 (2007). https://doi.org/10.1038/nprot.2007.446
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DOI: https://doi.org/10.1038/nprot.2007.446
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