Optical Tweezers.PPT

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Optical Tweezers.PPT

Optical Tweezers Sarah Nichols March 17, 2004 Outline Why use optical tweezers? Can manipulate living cells and organelles Can measure stretching of large molecules such as DNA Measure and manipulate mechanoenzymes and other physically acting molecules Relatively cheap and easy to build a simple setup Useful tool for learning various optics procedures and terms How it works dobjectdbeam - ray optics approximation is valid Reflection and refraction create opposing forces on a dielectric object If the beam is strong enough, the gradient force can overcome the scattering and gravitational forces Rays from edge of lens are deflected most, so they have the biggest trapping impact Components 20 mW diode laser 2 cylindrical lenses for shaping Mirrors for redirection 2 spherical lenses for resizing Periscope to raise the beam Dichroic mirror to direct beam into microscope Microscope with 100x oil immersion objective, eyepiece removed Camera connected to TV screen for viewing Layout Layout Optimization Beam must be aligned to travel exactly vertically through microscope For best trapping, beam should be spherical Optimal beam size overfills the objective slightly FWHM 6 mm for our setup (Amendola, 2001) Microscope objective does not collect light from infinity, so a lens should focus light 160 mm behind the objective Trapping most easily achieved with moving objects; surfactants used in bead solutions Results Despite numerous alignments, beam was weak until very recently Objective replacement improved beam strength Weak trapping demonstrated for yeast cells (~ 5 um) as well as 5 and 10 um polystyrene spheres With small yeast cells, can drag horizontally and vertically Results Questions Why is it easier to trap with yeast cells for a given size? (they exhibit more Brownian motion, but why?) Why does overfilling rule not hold? “Optimal” FWHM size: 6 mm. Measured size: 6-6.5 mm. Trapping size: 4 mm. Why does non-infinity correction make trapping harder? Suggested Improv

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