Abstract

Line-scanning microscopy is a technique with ability to deliver images with an higher acquisition rate than confocal microscopy. But it is accomplished at expense of the degradation of resolution for details parallel to sensor if slit detectors are used. With a linear image sensor it is possible to attenuate or even cancel this effect through the use of information stored in each pixel / light distribution across line pixels of the sensor. In spite of its great potential the use of linear image sensors and in particular the development of three-dimensional (3D) reconstruction methods that take into account its specificity is scarce. This led to our motivation to build a laboratory prototype of a bench stage-scanning microscope using a linear image sensor. We aim at improving lateral resolution isotropy but also image visualization and 3D mesh reconstruction using different optical setups particularly illumination modes, e.g., widefield and line-illumination. The versatility of the laboratory prototype namely its software for image acquisition, processing and visualization is important to attain this goal in the sense that it provides excellent means to develop and test algorithms. Several algorithms for 3D reconstruction were developed and are presented and discussed in this paper. Results of the application of these 3D reconstruction methods show the improvements on lateral resolution isotropy and depth discrimination achieved using algorithms integrating sensor geometry or spatial sampling rate. Also it is evidenced the impact of an insufficient spatial sampling rate from 3D mesh reconstructions.

© 2013 SPIE