Chen Bingying, Chakraborty Tonmoy, Daetwyler Stephan, Manton James D, Dean Kevin, Fiolka Reto
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
These authors contributed equally to this work.
Biomed Opt Express. 2020 Jun 19;11(7):3830-3842. doi: 10.1364/BOE.393931. eCollection 2020 Jul 1.
We present a beam splitter mask that can be easily added to a multiphoton raster scanning microscope to extend the depth of focus five-fold at a small loss in lateral resolution. The method is designed for ultrafast laser pulses or other light-sources featuring a low coherence length. In contrast to other methods of focus extension, our approach uniquely combines low complexity, high light-throughput and multicolor capability. We characterize the point spread function in a two-photon microscope and demonstrate fluorescence imaging of GFP labeled neurons in fixed brain samples as imaged with conventional and extended depth of focus two-photon microscopy.
我们展示了一种分束器掩膜，它可以轻松添加到多光子光栅扫描显微镜中，以将焦深扩展五倍，同时横向分辨率仅有小幅损失。该方法适用于超快激光脉冲或其他具有低相干长度的光源。与其他焦深扩展方法不同，我们的方法独特地结合了低复杂度、高透光率和多色能力。我们在双光子显微镜中表征了点扩散函数，并展示了在固定脑样本中对绿色荧光蛋白标记神经元进行的荧光成像，分别采用传统双光子显微镜和扩展焦深双光子显微镜成像。
