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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Gao, Ying Zhang, Yongli Sirinakis, George Ren, Yuxuan Xi, Zhiqun |
| Description | Country affiliation: United States Author Affiliation: Sirinakis G ( Department of Cell Biology, Yale University School of Medicine, 333 Cedar St., New Haven, Connecticut 06520, USA.) |
| Abstract | Optical trapping and single-molecule fluorescence are two major single-molecule approaches. Their combination has begun to show greater capability to study more complex systems than either method alone, but met many fundamental and technical challenges. We built an instrument that combines base-pair resolution dual-trap optical tweezers with single-molecule fluorescence microscopy. The instrument has complementary design and functionalities compared with similar microscopes previously described. The optical tweezers can be operated in constant force mode for easy data interpretation or in variable force mode for maximum spatiotemporal resolution. The single-molecule fluorescence detection can be implemented in either wide-field or confocal imaging configuration. To demonstrate the capabilities of the new instrument, we imaged a single stretched λ DNA molecule and investigated the dynamics of a DNA hairpin molecule in the presence of fluorophore-labeled complementary oligonucleotide. We simultaneously observed changes in the fluorescence signal and pauses in fast extension hopping of the hairpin due to association and dissociation of individual oligonucleotides. The combined versatile microscopy allows for greater flexibility to study molecular machines or assemblies at a single-molecule level. |
| ISSN | 00346748 |
| e-ISSN | 10897623 |
| Journal | Review of Scientific Instruments |
| Issue Number | 9 |
| Volume Number | 83 |
| Language | English |
| Publisher | American Institute of Physics |
| Publisher Date | 2012-09-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Microscopy, Fluorescence Instrumentation Optical Tweezers Bacteriophage Lambda Carbocyanines Metabolism Dna, Viral Genetics Inverted Repeat Sequences Research Support, N.i.h., Extramural Discipline Physics Discipline Chemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Instrumentation |
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