Single-Molecule Fluorescence Spectroscopy of Phase-Separated 10,12-Pentacosadynoic Acid Films

2021 ◽  
Author(s):  
Alfred Yeboah ◽  
David Sowah-Kuma ◽  
Wei Bu ◽  
Matthew F. Paige
Keyword(s):  
Fluorescence Spectroscopy ◽  
Single Molecule ◽  
Single Molecule Fluorescence ◽  
Single Molecule Fluorescence Spectroscopy

Combining Optical Trapping and Single-Molecule Fluorescence Spectroscopy:  Enhanced Photobleaching of Fluorophores

2004 ◽  
Vol 108 (20) ◽  
pp. 6479-6484 ◽  
Author(s):  
Meindert A. van Dijk ◽  
Lukas C. Kapitein ◽  
Joost van Mameren ◽  
Christoph F. Schmidt ◽  
Erwin J. G. Peterman
Keyword(s):  
Fluorescence Spectroscopy ◽  
Single Molecule ◽  
Optical Trapping ◽  
Single Molecule Fluorescence ◽  
Single Molecule Fluorescence Spectroscopy

scholarly journals Real-Time 3D Single Particle Tracking: Towards Active Feedback Single Molecule Spectroscopy in Live Cells

Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2826 ◽  
Author(s):  
Shangguo Hou ◽  
Courtney Johnson ◽  
Kevin Welsher
Keyword(s):  
Fluorescence Spectroscopy ◽  
Real Time ◽  
Temporal Resolution ◽  
Single Molecule ◽  
Particle Tracking ◽  
Single Particle ◽  
Single Particle Tracking ◽  
Single Molecule Fluorescence ◽  
Single Molecule Fluorescence Spectroscopy ◽  
Active Feedback

Single molecule fluorescence spectroscopy has been largely implemented using methods which require tethering of molecules to a substrate in order to make high temporal resolution measurements. However, the act of tethering a molecule requires that the molecule be removed from its environment. This is especially perturbative when measuring biomolecules such as enzymes, which may rely on the non-equilibrium and crowded cellular environment for normal function. A method which may be able to un-tether single molecule fluorescence spectroscopy is real-time 3D single particle tracking (RT-3D-SPT). RT-3D-SPT uses active feedback to effectively lock-on to freely diffusing particles so they can be measured continuously with up to photon-limited temporal resolution over large axial ranges. This review gives an overview of the various active feedback 3D single particle tracking methods, highlighting specialized detection and excitation schemes which enable high-speed real-time tracking. Furthermore, the combination of these active feedback methods with simultaneous live-cell imaging is discussed. Finally, the successes in real-time 3D single molecule tracking (RT-3D-SMT) thus far and the roadmap going forward for this promising family of techniques are discussed.

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