Pdf Construction And Calibration Of An Optical Trap On A Fluorescence Optical Microscope
File Name: construction and calibration of an optical trap on a fluorescence optical microscope.zip
These laser-based tweezers, or traps, have been employed in numerous biological experiments.
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This design is based on a conventional, inverted microscope, operating under plain bright field illumination. A single laser beam enables standard optical trapping and the measurement of molecular displacements and forces, whereas digital image processing affords real-time sample visualization with reduced noise and enhanced contrast. We have tested our trapping and imaging instrument by measuring the persistence length of individual double-stranded DNA molecules, and by following the stepping of single kinesin motor proteins along clearly imaged microtubules. The approach presented here provides a straightforward alternative for studies of biomaterials and individual biomolecules. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist.
Since the time of their introduction, optical tweezers OTs have grown to be a powerful tool in the hands of biologists. OTs use highly focused laser light to guide, manipulate, or sort target objects, typically in the nanoscale to microscale range. OTs have been particularly useful in making quantitative measurements of forces acting in cellular systems; they can reach inside living cells and be used to study the mechanical properties of the fluids and structures that they contain. As all the measurements are conducted without physically contacting the system under study, they also avoid complications related to contamination and tissue damage. From the manipulation of fluorescent nanodiamonds to chromosomes, cells, and free-swimming bacteria, OTs have now been extended to challenging biological systems such as the vestibular system in zebrafish. Here, we will give an overview of OTs, the complications that arise in carrying out OTs in vivo , and specific OT methods that have been used to address a range of otherwise inaccessible biological questions.
By setting up a well-calibrated single-beam optical trap within a fluorescence gradient force trap (optical tweezers) on an inverted fluorescence microscope. Three-axis translation stage with manual x–y–z micropositioners.
Modular Optical Tweezers
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