Fluorescent stereo microscopy for 3D surface profilometry and deformation mapping



Journal Title

Journal ISSN

Volume Title




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.;



Surface topography, Three dimensional sensing, Fluorescence microscopy, Three dimensional microscopy, Biomedical materials


"We acknowledge the support of DOE NEUP 09-818. We also thank the additional support from NSF DMR-0907291, CMMI-1031829, CMMI-1132174, and ECCS-1307997, and NIH R01 EB013212 and R01 DC011585. We appreciate Dr. Munehiro Asally and Dr. Gürol M. Süel at the University of California, San Diego for providing the biofilm samples."


©2013 Optical Society of America


Hu, Zhenxing, Huiyang Luo, Yingjie Du, and Hongbing Lu. 2013. "Fluorescent stereo microscopy for 3D surface profilometry and deformation mapping." Optics Express 21(10): 11808-11818