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- W4387520106 abstract "Techniques combining optical tweezers with fluorescence microscopy have become increasingly popular. Unfortunately, the high-power, infrared lasers used to create optical traps can have a deleterious effect on dye stability. Previous studies have shown that dye photobleaching is enhanced by absorption of visible fluorescence excitation plus infrared trap photons, a process that can be significantly reduced by minimizing simultaneous exposure to both light sources. Here, we report another photobleaching pathway that results from direct excitation by the trapping laser alone. Our results show that this trap-induced fluorescence loss is a two-photon absorption process, as demonstrated by a quadratic dependence on the intensity of the trapping laser. We further show that under conditions typical of many trap-based experiments, fluorescence emission of certain fluorophores near the trap focus can drop by 90% within one minute. We investigate how photostability is affected by the choice of dye molecule, excitation and emission wavelength, and labeled molecule. Finally, we discuss the different photobleaching pathways in combined trap-fluorescence measurements, which guide the selection of optimal dyes and conditions for more robust experimental protocols." @default.
- W4387520106 created "2023-10-12" @default.
- W4387520106 creator A5068744294 @default.
- W4387520106 creator A5086961765 @default.
- W4387520106 date "2023-10-01" @default.
- W4387520106 modified "2023-10-14" @default.
- W4387520106 title "Optical traps induce fluorophore photobleaching by two-photon excitation" @default.
- W4387520106 doi "https://doi.org/10.1016/j.bpj.2023.10.006" @default.
- W4387520106 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/37828742" @default.
- W4387520106 hasPublicationYear "2023" @default.
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