Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging

Kiryl D. Piatkevich, Ho Jun Suk, Suhasa B. Kodandaramaiah, Fumiaki Yoshida, Ellen M. DeGennaro, Mikhail Drobizhev, Thomas E. Hughes, Robert Desimone, Edward S. Boyden, Vladislav V. Verkhusha

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Abstract

Several series of near-infrared (NIR) fluorescent proteins (FPs) were recently engineered from bacterial phytochromes but were not systematically compared in neurons. To fluoresce, NIR FPs utilize an enzymatic derivative of heme, the linear tetrapyrrole biliverdin, as a chromophore whose level in neurons is poorly studied. Here, we evaluated NIR FPs of the iRFP protein family, which were reported to be the brightest in non-neuronal mammalian cells, in primary neuronal culture, in brain slices of mouse and monkey, and in mouse brain in vivo. We applied several fluorescence imaging modes, such as wide-field and confocal one-photon and two-photon microscopy, to compare photochemical and biophysical properties of various iRFPs. The iRFP682 and iRFP670 proteins exhibited the highest brightness and photostability under one-photon and two-photon excitation modes, respectively. All studied iRFPs exhibited efficient binding of the endogenous biliverdin chromophore in cultured neurons and in the mammalian brain and can be readily applied to neuroimaging.

Original languageEnglish (US)
Pages (from-to)2299-2309
Number of pages11
JournalBiophysical journal
Volume113
Issue number10
DOIs
StatePublished - Nov 21 2017

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ASJC Scopus subject areas

  • Biophysics

Cite this

Piatkevich, K. D., Suk, H. J., Kodandaramaiah, S. B., Yoshida, F., DeGennaro, E. M., Drobizhev, M., Hughes, T. E., Desimone, R., Boyden, E. S., & Verkhusha, V. V. (2017). Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging. Biophysical journal, 113(10), 2299-2309. https://doi.org/10.1016/j.bpj.2017.09.007