Hu Zhenxing, Luo Huiyang, Du Yingjie, Lu Hongbing
Department of Mechanical Engineering, 800 W Campbell Rd, the University of Texas at Dallas, Richardson, TX 75252, USA.
Opt Express. 2013 May 20;21(10):11808-18. doi: 10.1364/OE.21.011808.
Recently, mechanobiology has received increased attention. For investigation of biofilm and cellular tissue, measurements of the surface topography and deformation in real-time are a pre-requisite for understanding the growth mechanisms. In this paper, a novel three-dimensional (3D) fluorescent microscopic method for surface profilometry and deformation measurements is developed. In this technique a pair of cameras are connected to a binocular fluorescent microscope to acquire micrographs from two different viewing angles of a sample surface doped or sprayed with fluorescent microparticles. Digital image correlation technique is used to search for matching points in the pairing fluorescence micrographs. After calibration of the system, the 3D surface topography is reconstructed from the pair of planar images. When the deformed surface topography is compared with undeformed topography using fluorescent microparticles for movement tracking of individual material points, the full field deformation of the surface is determined. The technique is demonstrated on topography measurement of a biofilm, and also on surface deformation measurement of the biofilm during growth. The use of 3D imaging of the fluorescent microparticles eliminates the formation of bright parts in an image caused by specular reflections. The technique is appropriate for non-contact, full-field and real-time 3D surface profilometry and deformation measurements of materials and structures at the microscale.
近年来，力学生物学受到了越来越多的关注。对于生物膜和细胞组织的研究，实时测量表面形貌和变形是理解其生长机制的先决条件。本文开发了一种用于表面轮廓测量和变形测量的新型三维（3D）荧光显微镜方法。在该技术中，一对相机连接到双目荧光显微镜，以从掺杂或喷涂有荧光微粒的样品表面的两个不同视角获取显微图像。数字图像相关技术用于在配对的荧光显微图像中寻找匹配点。系统校准后，从这对平面图像重建三维表面形貌。当使用荧光微粒对单个材料点进行运动跟踪，将变形后的表面形貌与未变形的形貌进行比较时，即可确定表面的全场变形。该技术在生物膜的形貌测量以及生物膜生长过程中的表面变形测量中得到了验证。荧光微粒的三维成像消除了由镜面反射引起的图像中亮部的形成。该技术适用于微米尺度材料和结构的非接触、全场和实时三维表面轮廓测量及变形测量。
