Optimisation of FISH Methodology for the Study of 3D Structure of Cell Nuclei Using Micro Axial Tomography

• Authors: SKALNÍKOVÁ Magdalena; KOZUBEK Michal; MATULA Petr; BÁRTOVÁ Eva; EIPEL Heinz; HAUSMANN Michael
• Year of publication: 2002
• Type: Article in Proceedings
• Conference: Biophysics of the Genome and Its Interactions
• MU Faculty or unit: Faculty of Informatics
• Field: Biophysics
• Keywords: Chromatin structure; FISH; Micro axial tomography; High-resolution cytometry; fibre glass
• Description: Micro axial tomography is a new method for microscopic image acquisition of fluorescence in situ hybridisation (FISH) stained cell nuclei with improved spatial resolution. Automated axial tomography has grown up from a common Brno-Heidelberg collaboration of two scientific groups, which are interested in the study of higher order chromatin structure in FISH stained cells using epifluorescence and confocal laser scanning fluorescence microscopy. The aim of the common Brno-Heidelberg project is to combine high-resolution cytometry (HRCM) and 2đ-tilting device in order to obtain statistically significant data about the structure of cell nuclei using advanced microscopy and fast automated computer image acquisition and analysis. In this study a method for the preparation of cell nuclei attached to glass fibres has been developed. The optimised experimental procedure for the first time provides imaging of three-dimensionally conserved, FISH-stained cell nuclei fixed to a glass fibre

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