Construction of a High Resolution Microscope with Conventional and Holographic Optical Trapping Capabilities

Abstract

High resolution microscope systems with optical traps allow for precise manipulation of various refractive objects, such as dielectric beads 1 or cellular organelles 2,3 , as well as for high spatial and temporal resolution readout of their position relative to the center of the trap. The system described herein has one such "traditional" trap operating at 980 nm. It additionally provides a second optical trapping system that uses a commercially available holographic package to simultaneously create and manipulate complex trapping patterns in the field of view of the microscope 4,5 at a wavelength of 1,064 nm. The combination of the two systems allows for the manipulation of multiple refractive objects at the same time while simultaneously conducting high speed and high resolution measurements of motion and force production at nanometer and piconewton scale. Video LinkThe video component of this article can be found at

Cargo Transport at Microtubule Crossings: Evidence for Prolonged Tug-of-War between Kinesin Motors

“…3 B). Cargos were not slowing down due to high number of motors working together (12) or motor crowding (10,11), since motor velocities appeared healthy away from MT crossings (Fig. S2).…”

“…Measurements were performed as previously described (10). Briefly, beads and MTs were viewed using a Nikon Eclipse Ti microscope equipped with a high-magnification, high-NA objective (Nikon Plan Apo VC 100Â oil, 1.40 NA).…”

Single-molecule inhibition of human kinesin by adociasulfate-13 and -14 from the spongeCladocroce aculeata

“…Optical trapping experiments were performed using a previously described measurement setup (27). Briefly, beads and MTs were viewed using an inverted DIC microscope (Nikon Eclipse Ti-U) equipped with a high-magnification, high-NA objective (Nikon Plan Apo VC 100× oil, 1.40 NA).…”

Custom complex 3D microtubule networks for experimentation and engineering

“…Indeed, a few years ago we reported the construction of a novel instrument designed specifically for these types of experiments. 6 We combined two well-established technologies: holographic optical trapping and ordinary optical trapping with a capability for fast repositioning in the field of view in one experimental setup.…”