Photo‐isomerization of the Cyanine Dye Alexa‐Fluor 647 (AF‐647) in the Context of dSTORM Super‐Resolution Microscopy

Joshua Karlsson, Alex Laude, Michael J. Hall, Anthony Harriman

Tutkimustuotos: ArtikkeliTieteellinenvertaisarvioitu

11 Sitaatiot (Scopus)

Abstrakti

Cyanine dyes, as used in super-resolution fluorescence microscopy, undergo light-induced “blinking”, enabling localization of fluorophores with spatial resolution beyond the optical diffraction limit. Despite a plethora of studies, the molecular origins of this blinking are not well understood. Here, we examine the photophysical properties of a bio-conjugate cyanine dye (AF-647), used extensively in dSTORM imaging. In the absence of a potent sacrificial reductant, light-induced electron transfer and intermediates formed via the metastable, triplet excited state are considered unlikely to play a significant role in the blinking events. Instead, it is found that, under conditions appropriate to dSTORM microscopy, AF-647 undergoes reversible photo-induced isomerization to at least two long-lived dark species. These photo-isomers are characterized spectroscopically and their interconversion probed by computational means. The first-formed isomer is light sensitive and transforms to a longer-lived species in modest yield that could be involved in dSTORM related blinking. Permanent photobleaching of AF-647 occurs with very low quantum yield and is partially suppressed by the anaerobic redox buffer.
AlkuperäiskieliEnglanti
Sivut14983-14998
JulkaisuChemistry - A European Journal
DOI - pysyväislinkit
TilaJulkaistu - 13 syysk. 2019
Julkaistu ulkoisestiKyllä
OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

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