TY - JOUR
T1 - Near-Infrared Fluorescent Proteins Engineered from Bacterial Phytochromes in Neuroimaging
AU - Piatkevich, Kiryl D.
AU - Suk, Ho Jun
AU - Kodandaramaiah, Suhasa B.
AU - Yoshida, Fumiaki
AU - DeGennaro, Ellen M.
AU - Drobizhev, Mikhail
AU - Hughes, Thomas E.
AU - Desimone, Robert
AU - Boyden, Edward S.
AU - Verkhusha, Vladislav V.
N1 - Publisher Copyright:
© 2017 Biophysical Society
PY - 2017/11/21
Y1 - 2017/11/21
N2 - 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.
AB - 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.
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U2 - 10.1016/j.bpj.2017.09.007
DO - 10.1016/j.bpj.2017.09.007
M3 - Article
C2 - 29017728
AN - SCOPUS:85030651185
SN - 0006-3495
VL - 113
SP - 2299
EP - 2309
JO - Biophysical journal
JF - Biophysical journal
IS - 10
ER -