Exploratory Study on Light-Sheet Based Three-Dimensional Surface Topography

Light-sheet microscopy has attracted considerable attention because it is a fluorescence imaging technique with rapid optical sectioning capability for transparent samples. In this study, we report a new application based on light-sheet microscopy for exploratory investigation of three-dimensional surface topography of opaque objects. Instead of using inelastic scattering fluorescent signals, our method utilizes the elastic scattering of light from the surface of opaque samples, which are illuminated by a light sheet generated by a cylindrical lens. Through a simple structural modification by removing the fluorescent filter, the orthogonal imaging module can capture the elastically-scattered image. As the opaque object is scanned by a motorized stage, the light-sheet microscope acquires a series of sectional images, which can be stitched into a three-dimensional surface topography image. This method also offers the opportunity to visualize a 3D fingerprint at micron-level resolution. Therefore, this technique may be used in industry and the biomedical field for the measurement of surface microstructure. To our best knowledge, this is the first time a light-sheet microscopy is utilized to perform surface topography measurement.