Optical trapping is a powerful and sensitive technique that enables the direct measurement and manipulation of biological samples on the piconewton and nanometer scales. In examining the evolution and maturation of optical trapping instrumentation in the field of single-molecule biophysics, two main thrusts become apparent; a subset of instruments have aimed to push the limits of resolution, seeking to capture the finest details of biomechanical processes, while a second subset of instruments have explored the integration of optical trapping with other measurement techniques, with the goal of creating multidimensional, hybrid instruments. Unfortunately, many hybrid trapping instruments are forced to sacrifice measurement resolution, or relinquish their measurement capabilities entirely, when the combination of techniques cannot accommodate the stringent demands of traditional optical trapping detection methods. Thus, the full potential of hybrid optical trapping instruments has remained elusive. We have taken on this challenge by developing a highly-versatile optical tweezers instrument that can achieve sub-nanometer position resolution without the use of a second microscope objective, making high-resolution optical trapping measurements uniquely compatible with a broad array of bulky and/or opaque sample-side devices. Our instrument consists of a time-shared, dual optical trap implemented in an entirely custom-built, free-standing fluorescence microscope, with camera-based position detection. We have developed a robust and accurate set of tracking techniques that allow us to make the first direct measurements of relative bead displacement within an optical trap with sub-nanometer resolution using image tracking. These capabilities pave the way for the next generation of optical tweezer hybrids, which we envision as versatile tools that can integrate multiple measurement modalities and complex substrates without sacrificing resolution, to expand the breadth and clarity of our window into the microscopic and sub-microscopic biological worlds.
|Advisor:||Wang, Michelle D.|
|Commitee:||Arias, Tomas A., Vengalattore, Mukund M.|
|School Location:||United States -- New York|
|Source:||DAI-B 78/06(E), Dissertation Abstracts International|
|Keywords:||Dual trap, Image tracking, Optical trap, Optical tweezers, Resolution, Single molecule|
Copyright in each Dissertation and Thesis is retained by the author. All Rights Reserved
The supplemental file or files you are about to download were provided to ProQuest by the author as part of a
dissertation or thesis. The supplemental files are provided "AS IS" without warranty. ProQuest is not responsible for the
content, format or impact on the supplemental file(s) on our system. in some cases, the file type may be unknown or
may be a .exe file. We recommend caution as you open such files.
Copyright of the original materials contained in the supplemental file is retained by the author and your access to the
supplemental files is subject to the ProQuest Terms and Conditions of use.
Depending on the size of the file(s) you are downloading, the system may take some time to download them. Please be