Dynamic excitation for photophysical control and enhanced FRET sensing in an Anti-Brownian ELectrokinetic (ABEL) trap

Ayesha Ejaz; Kavya Vaidya; Allison H. Squires

doi:10.1117/12.2680788

5 October 2023 Dynamic excitation for photophysical control and enhanced FRET sensing in an Anti-Brownian ELectrokinetic (ABEL) trap

Ayesha Ejaz, Kavya Vaidya, Allison H. Squires

Author Affiliations +

Proceedings Volume 12649, Optical Trapping and Optical Micromanipulation XX; 126490W (2023) https://doi.org/10.1117/12.2680788
Event: SPIE Nanoscience + Engineering, 2023, San Diego, California, United States

Abstract

The Anti-Brownian ELectrokinetic (ABEL) trap uses electrophoresis or electroosmosis to trap fluorescent single molecules in free solution. Several photophysical parameters, such as brightness, lifetime, anisotropy, emission spectrum, can be simultaneously measured with very high precision. Here we demonstrate the added capabilities of the ABEL trap upon modifying the excitation technique. First, we utilize dynamic excitation brightness to observe photophysically induced responses in a trapped light-harvesting protein-pigment complex from cyanobacteria. Second, we trap fluorescently labeled biomolecules using two colors via pulsed interleaved excitation (PIE) to enable acceptor-corrected FRET measurements. These advances in excitation patterning for the ABEL trap allow for novel sensing abilities that combine the benefits of cutting-edge single-molecule imaging and spectroscopy with the long isotropic view of single molecules provided by the ABEL trap.

Citation Download Citation

Ayesha Ejaz, Kavya Vaidya, and Allison H. Squires "Dynamic excitation for photophysical control and enhanced FRET sensing in an Anti-Brownian ELectrokinetic (ABEL) trap", Proc. SPIE 12649, Optical Trapping and Optical Micromanipulation XX, 126490W (5 October 2023); https://doi.org/10.1117/12.2680788

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KEYWORDS

Fluorescence resonance energy transfer

Molecules

Dyes

Fluorescence

Photobleaching

Fluorescence intensity

Quantum efficiency

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