A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging

Janet Cusido, Sherif Shaban Ragab, Ek Raj Thapaliya, Subramani Swaminathan, Jaume Garcia-Amorós, M. Julia Roberti, Beatriz Araoz, Mercedes M A Mazza, Shiori Yamazaki, Amy M Scott, Francisco Raymo, Mariano L. Bossi

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically the ground-state absorption of the coumarin component. Selective excitation of the photochemical product then produces significant fluorescence and allows the detection of activated bioconjugates at the single-molecule level. Such fluorescence activation events can be exploited to resolve temporally individual emitters and reconstruct images of immunolabeled cells with subdiffraction resolution. Relying on a similar conjugation protocol, a model compound, incorporating the same chromophore of the photochemical product, can also be connected covalently to a secondary antibody. Stimulated emission can be exploited to deplete the excited state of the bioconjugated chromophores and switch their fluorescence off. These operating principles for fluorescence switching also permit the imaging of immunolabeled cells with subdiffraction resolution. Thus, these photoswitchable molecules, in combination with the labeling ability of antibodies, can evolve into valuable probes for bioimaging with superresolution.

Original languageEnglish (US)
Pages (from-to)12860-12870
Number of pages11
JournalJournal of Physical Chemistry C
Volume120
Issue number23
DOIs
StatePublished - Jun 16 2016

ASJC Scopus subject areas

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

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