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24 March 2005 Tissomics: two- and three-dimensional distribution of nuclei in brain tissue using laser scanning cytometry (LSC)
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Automated quantitative (i.e. stochiometric) analysis of tissues is of eminent importance in the understanding of all interactions between cells in their natural environment. In tissue cytometry a solid trigger is necessary in order to unequivocally differentiate between cellular and non-cellular events. This can be best performed by nuclear staining. Aim of this study was to analyze a brain tissue section by laser scanning cytometry (LSC) in order to depict the threedimensional distribution of nuclei in the tissue. To this end the section was measured in several foci and different nuclei detected in several depths of the tissue were assigned to the respective layer. Frozen sections of formalin-fixed rat or human brain tissue (120μm thickness) were incubated with propidiumiodide (PI) (50μg/ml) and covered on slides. For analysis by the LSC propidiumiodide was used as trigger. After a first analysis focussed on the top of the tissue, the focus was adjusted in 30μm steps deeper into the tissue. Per analysis data of at least 50,000 cells were acquired. After finishing measurements from all depths of the field were merged, i.e. data were combined into a composite data file. With the special features of the LSC it was possible to develop a method depicting the threedimensional distribution of the nuclei in solid tissue sections. LSC can be useful tool for this relatively new field of solid tissue cytometry termed tissomics. After evaluation of methods like this, so far not available data can be analysed for diagnostic purposes. By these studies we intend to demonstrate the power of the LSC for the routine pathological use. This should add up to the bright versatility of applications for the LSC as a cytometric instrument suitable for high throughput and high content analysis.
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Domnik Lenz, Anja Mittag, Birgit Mosch, Jozsef Bocsi, Thomas Arendt, and Attila Tarnok "Tissomics: two- and three-dimensional distribution of nuclei in brain tissue using laser scanning cytometry (LSC)", Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005);

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