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Abstract
A novel light-sheet microscopy (LSM) system that uses the laser triangulation method to quantitatively calculate and analyze the surface topography of opaque samples is discussed. A spatial resolution of at least 10 μm in z-direction, 10 μm in x-direction and 25 μm in y-direction with a large field-of-view (FOV) is achieved. A set of sample measurements that verify the system′s functionality in various applications are presented. The system has a simple mechanical structure, such that the spatial resolution is easily improved by replacement of the objective, and a linear calibration formula, which enables convenient system calibration. As implemented, the system has strong potential for, e.g., industrial sample line inspections, however, since the method utilizes reflected/scattered light, it also has the potential for three-dimensional analysis of translucent and layered structures.
Highlights
Micro-optical imaging technology is noninvasive, offers high speed and high resolution, and is used extensively in biological research as well as for industrial applications
Mechanical stylus measurements [4,5] have been used in recent years for 3D sample surface topography inspection, they damage the surface of the sample
We have previously proposed a light-sheet microscopy setup for qualitative 3D detection without distortion [27] by utilizing reflected or backscattered light
Summary
Micro-optical imaging technology is noninvasive, offers high speed and high resolution, and is used extensively in biological research as well as for industrial applications. Mechanical stylus measurements [4,5] have been used in recent years for 3D sample surface topography inspection, they damage the surface of the sample To avoid this drawback, several non-contact optical technologies that construct 3D spatial models through microscopic imaging have been proposed. Light-sheet microscopy (LSM) was first reported by Huisken et al [18] for in-vivo bio-imaging that demonstrated the method0 s high contrast and axial resolution, low photobleaching and low photo damage. It is worth mentioning that a few companies, such as Keyence or NEO Subsea AS, have launched commercial sensing systems using light-sheet technology for high-speed measurements of 3D surface topographies of non-transparent samples. The sample surface topography will modulate the light-sheet in the height direction and, based on this, we can construct a
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