Semi-automated FISH-stained interphase nuclei analysis in human genome studies
| Authors | |
|---|---|
| Year of publication | 1998 |
| Type | Article in Proceedings |
| Conference | Fluorescence Microscopy and Fluorescent Probes |
| MU Faculty or unit | |
| Citation | |
| Field | Use of computers, robotics and its application |
| Keywords | image analysis; automated microscopy; fluorescence in situ hybridization; interphase nuclei |
| Description | We have used fluorescence in situ hybridization (FISH) technique for the investigation of the arrangement of specific genes and chromosomes in human cells. Lymphocytes, bone marrow cells, human fibroblasts and other cell lines were studied. Results for normal and irradiated cells were compared in order to estimate the influence of radiation on the genes' arrangement. We compared also the difference in genes' arrangement between healthy donors and patients suffering from a specific disease related to a certain type of chromosomal aberration, such as BCR/ABL gene in the case of chronic myeloid leukaemia. Both epi-fluorescence and confocal microscopes were used for the observation. Both two-dimensional (2D) and three-dimensional (3D) analysis of interphase nuclei was performed. Besides, an algorithm for the estimation of 3D structure form 2D projections was proposed. The localization of interphase nuclei in the microscope was performed manually, image capturing and storage semi-automatically. Computer analysis was full-automatic with the possibility to analyse whole stack of images overnight. The results of the automatic analysis were verified visually on the computer monitor and corrected where necessary. The whole system is being rebuilt now into a full-automatic version based on Leica DMRXA motorized computer driven microscope. Algorithms for the image analysis such as nuclei and genes' segmentation as well as biological results of our studies are presented. |
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