Photoactivatable fluorophores for super-resolution imaging based on oxazine auxochromes

Erhan Deniz, Massimiliano Tomasulo, Janet Cusido, Ibrahim Yildiz, Marco Petriella, Mariano L. Bossi, Salvatore Sortino, Francisco Raymo

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

We synthesized five fluorophore-photochrome dyads designed to switch reversibly between nonfluorescent and fluorescent isomers under optical control. These compounds pair an oxazine photochrome to a biphenyl, fluorene, pyrene, coumarin, or cyanine fluorophore in their molecular skeleton and can be prepared in a single step from known precursors in yields ranging from 30 to 63%. Nuclear magnetic resonance spectroscopy indicates that the oxazine ring of these compounds opens and closes spontaneously on a millisecond time scale in acetonitrile at ambient temperature. Under these conditions, the fraction of ring-open isomer at equilibrium is negligible in all instances with the exception of the cyanine derivative, which instead is almost exclusively in this form. Absorption and emission spectroscopies demonstrate, however, that the fraction of ring-open isomer is sensitive to solvent polarity and increases with a transition from acetonitrile to methanol. Alternatively, the ring-open isomer can be populated photochemically or trapped with the addition of acid. In both instances, the characteristic absorption and emission bands of the 3H-indolium chromophores, embedded within the ring-open species, can clearly be observed in the visible region. In the case of the coumarin derivative, the brightness of this chromophoric fragment is sufficiently high to permit the imaging of individual molecules with excellent signal-to-noise ratios. In fact, the fluorescence of single fluorophore-photochrome dyads can be activated under the influence of ultraviolet inputs and the resulting species can be localized with nanoscale precision under visible illumination. Indeed, subdiffraction images of polymer nanoparticles, doped with this particular dyad, can be reconstructed with nanoscale resolution. Thus, our operating principles for fluorescence switching at the single-molecule level can offer the opportunity to overcome diffraction and, eventually, lead to the development of an entire family of probes for super-resolution fluorescence imaging.

Original languageEnglish
Pages (from-to)6058-6068
Number of pages11
JournalJournal of Physical Chemistry C
Volume116
Issue number10
DOIs
StatePublished - Mar 15 2012

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Oxazines
Fluorophores
Isomers
Imaging techniques
isomers
Fluorescence
rings
Acetonitrile
fluorescence
acetonitrile
Derivatives
Coumarins
Molecules
Pyrene
Emission spectroscopy
Chromophores
Absorption spectroscopy
optical control
Nuclear magnetic resonance spectroscopy
Methanol

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Electronic, Optical and Magnetic Materials
  • Surfaces, Coatings and Films
  • Energy(all)

Cite this

Photoactivatable fluorophores for super-resolution imaging based on oxazine auxochromes. / Deniz, Erhan; Tomasulo, Massimiliano; Cusido, Janet; Yildiz, Ibrahim; Petriella, Marco; Bossi, Mariano L.; Sortino, Salvatore; Raymo, Francisco.

In: Journal of Physical Chemistry C, Vol. 116, No. 10, 15.03.2012, p. 6058-6068.

Research output: Contribution to journalArticle

Deniz, E, Tomasulo, M, Cusido, J, Yildiz, I, Petriella, M, Bossi, ML, Sortino, S & Raymo, F 2012, 'Photoactivatable fluorophores for super-resolution imaging based on oxazine auxochromes', Journal of Physical Chemistry C, vol. 116, no. 10, pp. 6058-6068. https://doi.org/10.1021/jp211796p
Deniz E, Tomasulo M, Cusido J, Yildiz I, Petriella M, Bossi ML et al. Photoactivatable fluorophores for super-resolution imaging based on oxazine auxochromes. Journal of Physical Chemistry C. 2012 Mar 15;116(10):6058-6068. https://doi.org/10.1021/jp211796p
Deniz, Erhan ; Tomasulo, Massimiliano ; Cusido, Janet ; Yildiz, Ibrahim ; Petriella, Marco ; Bossi, Mariano L. ; Sortino, Salvatore ; Raymo, Francisco. / Photoactivatable fluorophores for super-resolution imaging based on oxazine auxochromes. In: Journal of Physical Chemistry C. 2012 ; Vol. 116, No. 10. pp. 6058-6068.
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