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Proceedings Paper

Acquisition of multiple image stacks with a confocal laser scanning microscope
Author(s): Werner Zuschratter; Thomas Steffen; Katharina Braun; Andreas Herzog; Bernd Michaelis; Henning Scheich
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Paper Abstract

Image acquisition at high magnification is inevitably correlated with a limited view over the entire tissue section. To overcome this limitation we designed software for multiple image-stack acquisition (3D-MISA) in confocal laser scanning microscopy (CLSM). The system consists of a 4 channel Leica CLSM equipped with a high resolution z- scanning stage mounted on a xy-monitorized stage. The 3D- MISA software is implemented into the microscope scanning software and uses the microscope settings for the movements of the xy-stage. It allows storage and recall of 70 xyz- positions and the automatic 3D-scanning of image arrays between selected xyz-coordinates. The number of images within one array is limited only by the amount of disk space or memory available. Although for most applications the accuracy of the xy-scanning stage is sufficient for a precise alignment of tiled views, the software provides the possibility of an adjustable overlap between two image stacks by shifting the moving steps of the xy-scanning stage. After scanning a tiled image gallery of the extended focus-images of each channel will be displayed on a graphic monitor. In addition, a tiled image gallery of individual focal planes can be created. In summary, the 3D-MISA allows 3D-image acquisition of coherent regions in combination with high resolution of single images.

Paper Details

Date Published: 9 June 1998
PDF: 10 pages
Proc. SPIE 3261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V, (9 June 1998); doi: 10.1117/12.310551
Show Author Affiliations
Werner Zuschratter, Federal Institute of Neurobiology (Germany)
Thomas Steffen, Federal Institute of Neurobiology (Germany)
Katharina Braun, Federal Institute of Neurobiology (Germany)
Andreas Herzog, Otto-von-Guericke Univ. Magdeburg (Germany)
Bernd Michaelis, Otto-von-Guericke Univ. Magdeburg (Germany)
Henning Scheich, Federal Institute of Neurobiology (Germany)

Published in SPIE Proceedings Vol. 3261:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing V
Thomas Taiwei Lu; Carol J. Cogswell; Jeremy M. Lerner; Jose-Angel Conchello; Jeremy M. Lerner; Thomas Taiwei Lu; Tony Wilson, Editor(s)

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