Abstract
Fluorescent protein-based biosensors are indispensable molecular tools for life science research. The invention and development of high-fidelity biosensors for a particular molecule or molecular event often catalyze important scientific breakthroughs. Understanding the structural and functional organization of brain activities remain a subject for which optical sensors are in desperate need and of growing interest. Here, we review genetically encoded fluorescent sensors for imaging neuronal activities with a focus on the design principles and optimizations of various sensors. New bioluminescent sensors useful for deep-tissue imaging are also discussed. By highlighting the protein engineering efforts and experimental applications of these sensors, we can consequently analyze factors influencing their performance. Finally, we remark on how future developments can fill technological gaps and lead to new discoveries.
Highlights
The human brain has an average of 86 billion neurons, forming complex neuronal networks that are essential for behavior, intelligence, learning, and memory [1]
Since circularly permuted YFP (cpYFP) have new N- and C-termini in close proximity to the chromophore [44], their use as FRET acceptors would result in different FRET performances, compared to their wild-type counterparts, due to an alteration of the relative orientation and dipole of the donor and acceptor chromophores
Further engineering of the Mg2+ - and Ca2+ -binding sites within the C-terminal lobe of troponin C (TnC), and use of alternative donor/acceptor pair (ECFP/Citrine cp174), improved the ion selectivity, dynamic range, chromophore [44], their use as FRET acceptors would result in different FRET performances, and response kinetics of TN-L15, in TN-XL
Summary
The human brain has an average of 86 billion neurons, forming complex neuronal networks that are essential for behavior, intelligence, learning, and memory [1]. Techniques are electrophysiology and optical neuroimaging, through which electric and neurochemical Complementary to these techniques are electrophysiology and optical neuroimaging, through signals in the brain can be detected and further related to neuronal and cortical functions [13]. The ‘gold standard’ for investigating measure electrical activity of cells with high sensitivity, the invasive requirement of physical contact neuronal functions [14,15], can directly measure electrical activity of cells with high sensitivity, the with tissuesrequirement and the accompanied poor spatial resolution its dominantpoor position neuroscience invasive of physical contact with tissues and lend the accompanied spatialinresolution to lend be challenged byposition the emerging optical neuroimaging techniques [16]. Unless otherwise specified in the main text, the reported value refers to a positive relationship between Ca2+ and fluorescence intensity or ratio
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