Here, we describe a method of acquisition of fast fluorescent signals with the help of the laser scanning confocal microscope (LSCM). Our method permits an increase in the temporal resolution of acquired signals. The method is based on LSCM recordings of fast fluorescent signals with the shortest achievable time sweep, which are performed with the help of a proprietary algorithm. A series of recordings is made in multiple steps; at each step, the fluorescent signal is incremented by a time interval smaller than the time sweep of the frame of LSCM. The size of the increment determines the achievable time resolution. The convolution of the recorded images results in a signal with the temporal resolution determined by the chosen time increment. This method was applied to register the change in fluorescence (calcium transient) of calcium dye preloaded into peripheral nerve endings by electrical stimulation of the motor nerve. Calculated parameters of the calcium transient were identical to the parameters obtained earlier with the help of a high-speed camera and photodiode. We conclude that the method described here can be applied for the registration of fast fluorescent signals by LSCM with a high spatial and temporal resolution.

中文翻译：

---

增强共聚焦显微镜获得的荧光信号的时间分辨率

在这里，我们描述了一种借助激光扫描共聚焦显微镜 (LSCM) 获取快速荧光信号的方法。我们的方法允许增加采集信号的时间分辨率。该方法基于具有最短可实现时间扫描的快速荧光信号的 LSCM 记录，这些记录是在专有算法的帮助下执行的。一系列记录是分多个步骤进行的；在每一步，荧光信号都以小于 LSCM 帧的时间扫描的时间间隔递增。增量的大小决定了可实现的时间分辨率。记录图像的卷积产生具有由所选时间增量确定的时间分辨率的信号。该方法用于记录通过电刺激运动神经预加载到周围神经末梢的钙染料的荧光（钙瞬变）变化。钙瞬变的计算参数与之前在高速相机和光电二极管的帮助下获得的参数相同。我们得出结论，此处描述的方法可应用于具有高空间和时间分辨率的 LSCM 对快速荧光信号的注册。