All-optical constant-force laser tweezers.
Optical tweezers are a powerful tool for the study of single biomolecules. Many applications require that a molecule be held under constant tension while its extension is measured. We present two schemes based on scanning-line optical tweezers to accomplish this, providing all-optical alternatives to force-clamp traps that rely on electronic feedback to maintain constant-force conditions for the molecule. In these schemes, a laser beam is rapidly scanned along a line in the focal plane of the microscope objective, effectively creating an extended one-dimensional optical potential over distances of up to 8 microm. A position-independent lateral force acting on a trapped particle is created by either modulating the laser beam intensity during the scan or by using an asymmetric beam profile in the back focal plane of the microscope objective. With these techniques, forces of up to 2.69 pN have been applied over distances of up to 3.4 microm with residual spring constants of <26.6 fN/microm. We used these techniques in conjunction with a fast position measurement scheme to study the relaxation of lambda-DNA molecules against a constant external force with submillisecond time resolution. We compare the results to predictions from the wormlike chain model.
Nambiar
Rajalakshmi
R
Department of Physics and Biophysics Research Division, University of Michigan, Ann Arbor, Michigan 48109-1120, USA.
Gajraj
Arivalagan
A
Meiners
Jens-Christian
JC
D055592:Biophysical Phenomena; D001703:Biophysics; D004247:DNA / Q000737:chemistry / Q000648:ultrastructure; D007834:Lasers; D008027:Light; D008846:Micromanipulation / Q000295:instrumentation* / Q000379:methods*; D008853:Microscopy / Q000379:methods; D008863:Microspheres; D015233:Models, Statistical; D012542:Scattering, Radiation; D013314:Stress, Mechanical; D013997:Time Factors
D004247:DNA