A High-throughput Image Correlation Method for Rapid Analysis of Fluorophore Photoblinking and Photobleaching Rates

Sehayek, Simon; Gidi, Yasser; Glembockyte, Viktorija; Brandão, Hugo B.; François, Paul; Cosa, Gonzalo; Wiseman, Paul W.

doi:10.1016/j.bpj.2019.11.1749

Cited by 1 publication

(2 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Relevance to Super-resolution Microscopy. The two photochemical events of most relevance to stochastic superresolution microscopy are irreversible photobleaching of the chromophore 97 and reversible light-induced blinking of the fluorescent species. 98 The triplet excited state, although formed in low quantum yield with all nonhalogenated cyanine dyes, has been implicated in the photobleaching mechanism.…”

Section: The Journal Ofmentioning

confidence: 99%

“…41,99 Recent laser flash photolysis studies have been interpreted in terms of the dication radical being a key intermediate, and the reported spectral characteristics are comparable to those described herein. 97 Our kinetic measurements indicate that Cy 2+• decays via a second-order process at very low dye concentrations, but the half-life is highly sensitive to the molecular structure (Table 2). At higher dye concentration, Cy 2+• undergoes bimolecular association with a ground-state dye molecule to form the corresponding dimer trication radical.…”

Section: The Journal Ofmentioning

confidence: 99%

See 1 more Smart Citation

Pulse Radiolysis Investigation of Radicals Derived from Water-Soluble Cyanine Dyes: Implications for Super-resolution Microscopy

2021

View full text Add to dashboard Cite

Light-induced blinking, an inherent feature of many forms of super-resolution microscopy, has been linked to transient reduction of the fluorescent cyanine dye used as an imaging agent. There is, however, only scant literature information related to one-electron reduced cyanine dyes, especially in an aqueous environment. Here, we examine a small series of cyanine dyes, possessing disparate π-conjugation lengths, under selective reducing or oxidizing conditions. The experiment allows recording of both differential absorption spectra and decay kinetics of the resultant one-electron reduced or oxidized transient species in water. Relative to the ground state, absorption transitions for the various radicals are weak and somewhat broadened but do allow correlation with the π-conjugation length. In all cases, absorption maxima lie to the blue of the main ground-state transition. Under anaerobic conditions, the transient species decay on the microsecond to millisecond time scale, with the mean lifetime depending on molecular structure, radiation dose, and dye concentration. The experimental absorption spectra recorded for the one-electron reduced radicals and the presumed dimer cation radical compare well to spectra obtained from time-dependent density functional theory calculations. The results allow conclusions to be drawn regarding the plausibility of the reduced species being responsible for light-induced blinking in direct stochastic optical reconstruction microscopy.

show abstract

Section: The Journal Ofmentioning

confidence: 99%