Sun Hongyue, Song Bing, Dong Hongpai, Reid Brian, Player Michael A, Watson John, Zhao Min
University of Aberdeen, School of Engineering, Aberdeen AB24 3UE, Scotland, United Kingdom.
J Biomed Opt. 2008 Jan-Feb;13(1):014007. doi: 10.1117/1.2841050.
Digital in-line holography offers some significant advantages over conventional optical holography and microscopy to image biological specimens. By combining holography with digital video microscopy, an in-line holographic video microscope is developed and is capable of recording spatial 3D holographic images of biological specimens, while preserving the time dimension. The system enables high-speed video recording of fast cell movement, such as the rapid movement of blood cells in the blood stream in vivo. This capability is demonstrated with observations of fast 3-D movement of live cells in suspension cultures in response to a gentle shake to the Petri dish. The experimental and numerical procedures are incorporated with a fast reconstruction algorithm for reconstruction of holographic video frames at various planes (z axis) from the hologram and along the time axis. The current system enables both lateral and longitudinal resolutions down to a few micrometers. Postreconstruction processing of background subtraction is utilized to eliminate noise caused by scattered light, thereby enabling visualization of, for example, blood streams of live Xenopos tadpoles. The combination of digital holography and microscopy offers unique advantages for imaging of fast moving cells and other biological particles in three dimensions in vivo with high spatial and temporal resolution.
与传统光学全息术和显微镜相比，数字同轴全息术在对生物标本成像方面具有一些显著优势。通过将全息术与数字视频显微镜相结合，开发出了一种同轴全息视频显微镜，它能够记录生物标本的空间三维全息图像，同时保留时间维度。该系统能够对快速的细胞运动进行高速视频记录，比如体内血流中血细胞的快速运动。通过观察悬浮培养的活细胞在培养皿受到轻轻摇晃时的快速三维运动，证明了这一能力。实验和数值程序结合了一种快速重建算法，用于从全息图沿时间轴在不同平面（z轴）重建全息视频帧。当前系统能够实现低至几微米的横向和纵向分辨率。利用重建后背景减法的后处理来消除散射光引起的噪声，从而能够观察例如活非洲爪蟾蝌蚪的血流。数字全息术和显微镜的结合为在体内以高空间和时间分辨率对快速移动的细胞和其他生物粒子进行三维成像提供了独特优势。
