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

Woei Ming Lee, Peter J. Reece, Robert F. Marchington, Nikolaus K. Metzger, Kishan Dholakia

Research output: Contribution to journalArticlepeer-review

81 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)3226-3238
Number of pages13
JournalNature Protocols
Volume2
Issue number12
DOIs
Publication statusPublished - 2007

Keywords

  • SINGLE-MOLECULE
  • DNA MECHANICS
  • TWEEZERS
  • PARTICLES
  • FORCES

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