High-Resolution Optical Tweezers Combined With Single-Molecule Confocal Microscopy.
Methods Enzymol
; 582: 137-169, 2017.
Article
en En
| MEDLINE
| ID: mdl-28062033
We describe the design, construction, and application of an instrument combining dual-trap, high-resolution optical tweezers and a confocal microscope. This hybrid instrument allows nanomechanical manipulation and measurement simultaneously with single-molecule fluorescence detection. We present the general design principles that overcome the challenges of maximizing optical trap resolution while maintaining single-molecule fluorescence sensitivity, and provide details on the construction and alignment of the instrument. This powerful new tool is just beginning to be applied to biological problems. We present step-by-step instructions on an application of this technique that highlights the instrument's capabilities, detecting conformational dynamics in a nucleic acid-processing enzyme.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ADN Helicasas
/
Microscopía Confocal
/
Pinzas Ópticas
/
Imagen Individual de Molécula
Idioma:
En
Revista:
Methods Enzymol
Año:
2017
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Estados Unidos