Construction and calibration of an optical trap on a fluorescence optical microscope.
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.
Lee
Woei Ming
WM
SUPA, School of Physics and Astronomy, University of St. Andrews, North Haugh, St. Andrews KY16 9SS, UK.
Reece
Peter J
PJ
Marchington
Robert F
RF
Metzger
Nikolaus K
NK
Dholakia
Kishan
K
D008846:Micromanipulation / Q000295:instrumentation; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D008863:Microspheres; D052898:Optical Tweezers; D055095:Optics and Photonics / Q000295:instrumentation; D010316:Particle Size