Construction and calibration of an optical trap on a fluorescence optical microscope

  • Lee W
  • Reece P
  • Marchington R
 et al. 
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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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Authors

  • Rob MarchingtonCambridge Consultants Ltd.

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  • Woei Ming Lee

  • Peter J. Reece

  • Nikolaus K. Metzger

  • Kishan Dholakia

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