Zhang Pengfei, Phipps Mary E, Goodwin Peter M, Werner James H
Washington University in St. Louis, Department of Biomedical Engineering, Optical Imaging Laboratory, One Brookings Drive, St. Louis, Missouri 63130-4899, United States.
Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, United States.
J Biomed Opt. 2016 Oct 1;21(10):100502. doi: 10.1117/1.JBO.21.10.100502.
We have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as many photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.
我们开发了一种光片显微镜，该显微镜使用双贝塞尔光束的共焦扫描进行照明。数字微镜器件（DMD）放置在用于收集荧光的物镜的中间像平面中，并被编程为两行像素处于“开”状态，使得DMD用作空间滤波器，以抑制由贝塞尔光束旁瓣产生的离焦背景。通过对人A431细胞中荧光标记的肌动蛋白进行成像，证明了该显微镜的光学切片和离焦背景抑制能力。双贝塞尔光束系统能够在每次成像扫描中检测到两倍数量的光子，这对于低光应用（例如单分子定位）或高速成像且具有优异信噪比的情况很有用。虽然该方法是针对两个贝塞尔光束进行演示的，但这种方法可扩展到更多光束。
