Photophysical properties of Cerulean and Venus fluorescent proteins

Pabak Sarkar, Srinagesh V. Koushik, Steven S. Vogel, Ignacy Gryczynski, Zygmunt Gryczynski

Research output: Contribution to journalArticlepeer-review

37 Scopus citations


Cerulean and Venus are recently developed fluorescent proteins, often used as a donor-acceptor pair by researchers in Förster resonance energy transfer-based colocalization studies. We characterized the fluorescent properties of these two proteins in a broad spectral range (form ultraviolet to visible region). Excitation spectra, lifetimes, and polarization spectra show significant energy transfer from aromatic amino acids to the fluorescent protein chromophore. High steady-state anisotropy values and the lack of a fast component in anisotropy decays show that the fluorescent protein chromophore is rigidly fixed within the protein structure. Furthermore, we show that the chromophores are not accessible to external quenchers, such as acrylamide or potassium iodide (KI), allowing the removal of "unwanted" background in the environment with external quencher, while leaving the Cerulean/Venus fluorescence unchanged.

Original languageEnglish
Article number034047
JournalJournal of Biomedical Optics
Issue number3
StatePublished - 2009


  • Förster resonance energy transfer
  • fluorescence anisotropy
  • fluorescence timeline


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