Duocastella Martí, Vicidomini Giuseppe, Diaspro Alberto
Opt Express. 2014 Aug 11;22(16):19293-301. doi: 10.1364/OE.22.019293.
Maximizing the amount of spatiotemporal information retrieved in confocal laser scanning microscopy is crucial to understand fundamental three-dimensional (3D) dynamic processes in life sciences. However, current 3D confocal microscopy is based on an inherently slow stepwise process that consists of acquiring multiple 2D sections at different focal planes by mechanical or optical z-focus translation. Here, we show that by using an acoustically-driven optofluidic lens integrated in a commercial confocal system we can capture an entire 3D image in a single step. Our method is based on continuous axial scanning at speeds as high as 140 kHz combined with fast readout. In this way, one or more focus sweeps are produced on a pixel by pixel basis and the detected photons can be assigned to their corresponding focal plane enabling simultaneous multiplane imaging. We exemplify this method by imaging calibration and biological fluorescence samples. These results open the door to exploring new fundamental processes in science with an unprecedented time resolution.
在共聚焦激光扫描显微镜中最大化时空信息的获取量对于理解生命科学中的基本三维（3D）动态过程至关重要。然而，当前的3D共聚焦显微镜基于一种固有的缓慢逐步过程，该过程包括通过机械或光学z轴聚焦平移在不同焦平面获取多个2D切片。在此，我们展示了通过在商业共聚焦系统中集成一个声学驱动的光流体透镜，我们可以在一步中捕获整个3D图像。我们的方法基于高达140 kHz的速度进行连续轴向扫描并结合快速读出。通过这种方式，逐像素地产生一个或多个聚焦扫描，并且检测到的光子可以被分配到其相应的焦平面，从而实现同时多平面成像。我们通过对校准和生物荧光样本成像来例证这种方法。这些结果为以前所未有的时间分辨率探索科学中的新基本过程打开了大门。
