Simultaneous multiplane confocal microscopy using acoustic tunable lenses
Simultaneous multiplane confocal microscopy using acoustic tunable lenses
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DOI：
10.1364/oe.22.019293
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发表时间：
2014-08-11
期刊：
影响因子：
3.8
通讯作者：
Diaspro, Alberto
中科院分区：
文献类型：
--
作者：
Duocastella, Marti;Vicidomini, Giuseppe;Diaspro, Alberto
最大限度地提高共聚焦激光扫描显微镜中检索的时空信息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，目前的3D共聚焦显微镜是基于固有的缓慢的逐步过程，该过程包括通过机械或光学z焦点平移在不同的焦平面处获取多个2D截面。在这里，我们表明，通过使用集成在商业共焦系统中的声驱动的光流透镜，我们可以在一个步骤中捕获整个3D图像。我们的方法是基于连续轴向扫描速度高达140 kHz结合快速读出。以这种方式，在逐像素的基础上产生一个或多个聚焦扫描，并且可以将探测到的光子分配给它们对应的焦平面，从而实现同时的多平面成像。我们通过成像校准和生物荧光样品验证了该方法。这些结果为以前所未有的时间分辨率探索科学中新的基本过程打开了大门。(C)2014年美国光学学会
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. (C) 2014 Optical Society of America