NeuroTechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
NeuroTechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Cell Rep. 2019 Jan 15;26(3):802-813.e4. doi: 10.1016/j.celrep.2018.12.088.
Imaging voltage using fluorescent-based sensors could be an ideal technique to probe neural circuits with high spatiotemporal resolution. However, due to insufficient signal-to-noise ratio (SNR), imaging membrane potential in mammalian preparations is still challenging. In recent years, many genetically encoded voltage indicators (GEVIs) have been developed. To compare them and guide decisions on which GEVI to use, we have characterized side by side the performance of eight GEVIs that represent different families of molecular constructs. We tested GEVIs in vitro with 1-photon imaging and in vivo with 1-photon wide-field imaging and 2-photon imaging. We find that QuasAr2 exhibited the best performance in vitro, whereas only ArcLight-MT could be used to reliably detect electrical activity in vivo with 2-photon excitation. No single GEVI was ideal for every experiment. These results provide a guide for choosing optimal GEVIs for specific applications.
使用基于荧光的传感器进行电压成像可能是一种用高时空分辨率探测神经回路的理想技术。然而，由于信噪比（SNR）不足，在哺乳动物制剂中成像膜电位仍然具有挑战性。近年来，已经开发出许多基因编码的电压指示剂（GEVI）。为了对它们进行比较，并指导选择要使用的 GEVI，我们并排表征了代表不同分子结构家族的八种 GEVI 的性能。我们用单光子成像在体外测试了 GEVIs，并在体内用单光子宽场成像和双光子成像进行了测试。我们发现 QuasAr2 在体外表现出最佳性能，而只有 ArcLight-MT 可以用双光子激发在体内可靠地检测到电活动。没有一个 GEVI 适用于每个实验。这些结果为选择特定应用的最佳 GEVI 提供了指导。
