Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium Indicator.
Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium Indicator.
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DOI：
10.3389/fcell.2022.880107
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发表时间：
2022
影响因子：
5.5
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中科院分区：
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近红外（NIR）基因编码钙指标（GECIs）正在成为神经科学研究的有力工具。由于其光谱特性，使用近红外geci有助于避免由于哺乳动物组织中身体色素吸收、光散射和自身荧光而造成的信号损失。此外，当与普通荧光指示器和光遗传学致动器一起使用时，近红外geci不会受到交叉激发伪影的影响。虽然已经开发了几种近红外GECI，但目前还没有一种近红外GECI能够将电池的高亮度和光稳定性与小尺寸和快速响应动力学相结合。在这里，我们报道了一个小的基于fret的近红外荧光钙指示剂iGECInano。我们在体外，在非神经元哺乳动物细胞和小鼠原代神经元中表征iGECInano。与其他近红外GECIs相比，iGECInano在信噪比和响应动力学方面有所改善。
Near-infrared (NIR) genetically encoded calcium indicators (GECIs) are becoming powerful tools for neuroscience. Because of their spectral characteristics, the use of NIR GECIs helps to avoid signal loss from the absorption by body pigments, light-scattering, and autofluorescence in mammalian tissues. In addition, NIR GECIs do not suffer from cross-excitation artifacts when used with common fluorescent indicators and optogenetics actuators. Although several NIR GECIs have been developed, there is no NIR GECI currently available that would combine the high brightness in cells and photostability with small size and fast response kinetics. Here, we report a small FRET-based NIR fluorescent calcium indicator iGECInano. We characterize iGECInano in vitro, in non-neuronal mammalian cells, and primary mouse neurons. iGECInano demonstrates the improvement in the signal-to-noise ratio and response kinetics compared to other NIR GECIs.