| Chapter title |
High-Resolution “Fleezers”: Dual-Trap Optical Tweezers Combined with Single-Molecule Fluorescence Detection
|
|---|---|
| Chapter number | 8 |
| Book title |
Optical Tweezers
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6421-5_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6419-2, 978-1-4939-6421-5
|
| Authors |
Kevin D. Whitley, Matthew J. Comstock, Yann R. Chemla, Whitley, Kevin D., Comstock, Matthew J., Chemla, Yann R. |
| Abstract |
Recent advances in optical tweezers have greatly expanded their measurement capabilities. A new generation of hybrid instrument that combines nanomechanical manipulation with fluorescence detection-fluorescence optical tweezers, or "fleezers"-is providing a powerful approach to study complex macromolecular dynamics. Here, we describe a combined high-resolution optical trap/confocal fluorescence microscope that can simultaneously detect sub-nanometer displacements, sub-piconewton forces, and single-molecule fluorescence signals. The primary technical challenge to these hybrid instruments is how to combine both measurement modalities without sacrificing the sensitivity of either one. We present general design principles to overcome this challenge and provide detailed, step-by-step instructions to implement them in the construction and alignment of the instrument. Lastly, we present a set of protocols to perform a simple, proof-of-principle experiment that highlights the instrument capabilities. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 54 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 15 | 28% |
| Researcher | 10 | 19% |
| Student > Bachelor | 5 | 9% |
| Student > Master | 5 | 9% |
| Student > Doctoral Student | 2 | 4% |
| Other | 7 | 13% |
| Unknown | 10 | 19% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 18 | 33% |
| Physics and Astronomy | 12 | 22% |
| Chemistry | 5 | 9% |
| Agricultural and Biological Sciences | 4 | 7% |
| Engineering | 3 | 6% |
| Other | 2 | 4% |
| Unknown | 10 | 19% |