A novel approach to evaluation of the spatial relationship of fluorescent signals in ImunnoFISH analysis of different kind.

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Authors

STEJSKAL Stanislav MATULA Pavel DANĚK Ondřej MAŠKA Martin KRONTORÁD KOUTNÁ Irena KOZUBEK Michal

Year of publication 2010
Type Article in Proceedings
Conference Advances in Molecular and Cancer Biology, 2010
MU Faculty or unit

Faculty of Informatics

Citation
Field Genetics and molecular biology
Keywords confocal microscopy; image analysis; fluorescent in situ hybridisation; immunoflourescence; nucleus
Description Confocal fluorescence microscopy enables the study and observation of the different structures in the cell in the high resolution. Its role in epigenetic research is finding spatial distribution and interaction of factors that can affect gene expression without change in genetic code. Combination of specific staining of DNA sequences and nuclear proteins (ImmunoFISH) help us to describe the protein distribution in the neighbourhood of targeted genes. In our early studies we had focused on the mutual spatial relation between genetic loci with described genetic activity and nuclear proteins commonly linked with the regulation of gene activity. Unfortunately, a broad distribution of the protein signal disabled the classical co-localisation analysis and we had to find an alternative approach for image analysis. Output of the presented image analysis is helpful for the co-localisation analysis of image data with broad distribution of signal and suitable for statistical analysis. All steps of analysis were implemented into the Acquiarium software.
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