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 journalArticle

71 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 forthe 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 Dec 13

Fingerprint

Optical Tweezers
Calibration
Microscopes
Fluorescence
Optical tweezers
Biological systems
Transducers
Microspheres
Macromolecules
Nanoparticles

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Lee, Woei Ming ; Reece, Peter J. ; Marchington, Robert F. ; Metzger, Nikolaus K. ; Dholakia, Kishan. / Construction and calibration of an optical trap on a fluorescence optical microscope. In: Nature Protocols. 2007 ; Vol. 2, No. 12. pp. 3226-3238.
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Construction and calibration of an optical trap on a fluorescence optical microscope. / Lee, Woei Ming; Reece, Peter J.; Marchington, Robert F.; Metzger, Nikolaus K.; Dholakia, Kishan.

In: Nature Protocols, Vol. 2, No. 12, 13.12.2007, p. 3226-3238.

Research output: Contribution to journalArticle

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