Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners
Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2004.
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| Format: | Thesis |
| Language: | en_US |
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Massachusetts Institute of Technology
2005
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| Online Access: | http://hdl.handle.net/1721.1/27059 |
| _version_ | 1826208624426352640 |
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| author | Ferrer, Jorge M., 1976- |
| author2 | Roger D. Kamm and Matthew J. Lang. |
| author_facet | Roger D. Kamm and Matthew J. Lang. Ferrer, Jorge M., 1976- |
| author_sort | Ferrer, Jorge M., 1976- |
| collection | MIT |
| description | Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2004. |
| first_indexed | 2024-09-23T14:08:35Z |
| format | Thesis |
| id | mit-1721.1/27059 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:08:35Z |
| publishDate | 2005 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/270592019-04-10T07:56:23Z Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners Ferrer, Jorge M., 1976- Roger D. Kamm and Matthew J. Lang. Massachusetts Institute of Technology. Dept. of Mechanical Engineering. Massachusetts Institute of Technology. Dept. of Mechanical Engineering. Mechanical Engineering. Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2004. Includes bibliographical references (p. 75-77). Actin filaments are a major constituent of the cytoskeleton in most eukaryotic cells. They function as a connection between the cell body to the focal adhesions in order to transmit forces into and out of the cell. During the force transduction process, many proteins bind to actin filaments in order to initiate a signaling cascade that reaches the cell nucleus. However, the effects of forces in the binding kinetics between actin filaments and actin binding proteins are unknown. This work proposes an experimental setup to study the force-dependent binding kinetics of such proteins at the single molecule level by using an instrument that combines optical trapping with single molecule fluorescence. The main focus of this work was the design and construction of the experimental equipment. The results show position detection capabilities with a resolution of 5 nm. Also, the trap stiffness recorded was in the order of 0.05 pN/nm. With the combination of position and trap stiffness, the force resolution of the instrument is about 0.25 pN. Also, a photobleaching event for a single dye molecule was recorded, proving the single molecule fluorescence capabilities. In addition, a complete experimental assay is described in order to perform studies on how force application affects the binding of actin and actin binding proteins. by Jorge M. Ferrer. S.M. 2005-09-06T21:31:57Z 2005-09-06T21:31:57Z 2004 2004 Thesis http://hdl.handle.net/1721.1/27059 56801935 en_US M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 77 p. 4413183 bytes 4421182 bytes application/pdf application/pdf application/pdf Massachusetts Institute of Technology |
| spellingShingle | Mechanical Engineering. Ferrer, Jorge M., 1976- Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title | Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title_full | Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title_fullStr | Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title_full_unstemmed | Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title_short | Combined optical trapping and single molecule fluorescence to study the force-dependent binding kinetics between filamentous actin and its partners |
| title_sort | combined optical trapping and single molecule fluorescence to study the force dependent binding kinetics between filamentous actin and its partners |
| topic | Mechanical Engineering. |
| url | http://hdl.handle.net/1721.1/27059 |
| work_keys_str_mv | AT ferrerjorgem1976 combinedopticaltrappingandsinglemoleculefluorescencetostudytheforcedependentbindingkineticsbetweenfilamentousactinanditspartners |