Difference between revisions of "Optical trap"
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==Setup and alignment== | ==Setup and alignment== | ||
===Remove the optics=== | ===Remove the optics=== | ||
| − | |||
===Collimating and adjusting the fiber port=== | ===Collimating and adjusting the fiber port=== | ||
| + | ===Initial laser alignment=== | ||
| + | ===Beam expander coarse adjustment=== | ||
| + | ===Condenser adjustment=== | ||
===Connecting the piezo stage=== | ===Connecting the piezo stage=== | ||
===Fine adjusting the beam expander=== | ===Fine adjusting the beam expander=== | ||
Revision as of 16:03, 26 June 2012
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Introduction
Optical tweezers can exert measurable forces on micron-scale dielectric particles. This capability offers a unique and valuable tool for manipulating and measuring cell components at the single molecule level. For example, optical traps have been used extensively to investigate the mechanical properties of biological polymers and the force generation mechanisms of molecular motors. In many studies, optical tweezers apply force to functionalized microspheres, which act as convenient handles attached to molecules of interest.
To make quantitative force measurements, the instrument records the displacement of a trapped microsphere over time. For small displacements, the exerted force is very nearly proportional to displacement, so the trap can be modeled as a linear spring. Accurate force and position measurements depend on careful calibration of the position detector responsivity, G, and the trap stiffness α, also called the spring constant. The stiffness is a function of trapping laser power, bead size, bead composition, and optical properties of the sample.
This page has tips for setting up and aligning an optical trap. It discusses three methods for obtaining the spring constant and two methods for measuring α.
Overview of the instrument
Setup and alignment
Remove the optics
Collimating and adjusting the fiber port
Initial laser alignment
Beam expander coarse adjustment
Condenser adjustment
Connecting the piezo stage
Fine adjusting the beam expander
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OTKB software
Starting the software
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Calibration
Measuring R by scanning a stuck bead
PSD method
Equipartition method
Stokes method
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