论文标题
神经元尖峰和阈值活性的宽视野荧光寿命成像
Wide-field fluorescence lifetime imaging of neuron spiking and sub-threshold activity in vivo
论文作者
论文摘要
电压敏感的荧光探针的发展表明荧光寿命是生物系统电活动的有希望的读数。现有方法无法实现神经科学应用中电压成像所需的速度和灵敏度。在这里，我们证明了宽场电荧光寿命成像（EO-FLIM）允许在KHz框架采集速率上进行终生成像，在空间上解决动作电位传播以及现场成人果蝇的亚阈值神经活动。单电压录音达到了1 kHz的终生分辨率$ <5 $ ps。终身读数受到光子射击噪声的限制，该方法可强烈拒绝运动伪像和技术噪声源。记录显示局部跨膜去极化，两种具有不同荧光寿命的尖峰以及将尖峰的相位锁定到外部机械刺激上。
The development of voltage-sensitive fluorescent probes suggests fluorescence lifetime as a promising readout for electrical activity in biological systems. Existing approaches fail to achieve the speed and sensitivity required for voltage imaging in neuroscience applications. Here we demonstrate that wide-field electro-optic fluorescence lifetime imaging (EO-FLIM) allows lifetime imaging at kHz frame acquisition rates, spatially resolving action potential propagation and sub-threshold neural activity in live adult Drosophila. Lifetime resolutions of $< 5$ ps at 1 kHz were achieved for single cell voltage recordings. Lifetime readout is limited by photon shot noise and the method provides strong rejection of motion artifacts and technical noise sources. Recordings revealed local transmembrane depolarizations, two types of spikes with distinct fluorescence lifetimes, and phase locking of spikes to an external mechanical stimulus.
