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IMACULAT — An Open Access Package for the Quantitative Analysis of Chromosome Localization in the Nucleus

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  • Ishita Mehta
  • Sandeep Chakraborty
  • Basuthkar J Rao

Abstract

The alteration in the location of the chromosomes within the nucleus upon action of internal or external stimuli has been implicated in altering genome function. The effect of stimuli at a whole genome level is studied by using two-dimensional fluorescence in situ hybridization (FISH) to delineate whole chromosome territories within a cell nucleus, followed by a quantitative analysis of the spatial distribution of the chromosome. However, to the best of our knowledge, open access software capable of quantifying spatial distribution of whole chromosomes within cell nucleus is not available. In the current work, we present a software package that computes localization of whole chromosomes - Image Analysis of Chromosomes for computing localization (IMACULAT). We partition the nucleus into concentric elliptical compartments of equal area and the variance in the quantity of any chromosome in these shells is used to determine its localization in the nucleus. The images are pre-processed to remove the smudges outside the cell boundary. Automation allows high throughput analysis for deriving statistics. Proliferating normal human dermal fibroblasts were subjected to standard a two-dimensional FISH to delineate territories for all human chromosomes. Approximately 100 images from each chromosome were analyzed using IMACULAT. The analysis corroborated that these chromosome territories have non-random gene density based organization within the interphase nuclei of human fibroblasts. The ImageMagick Perl API has been used for pre-processing the images. The source code is made available at www.sanchak.com/imaculat.html.

Suggested Citation

  • Ishita Mehta & Sandeep Chakraborty & Basuthkar J Rao, 2013. "IMACULAT — An Open Access Package for the Quantitative Analysis of Chromosome Localization in the Nucleus," PLOS ONE, Public Library of Science, vol. 8(4), pages 1-7, April.
    • Handle: RePEc:plo:pone00:0061386
    DOI: 10.1371/journal.pone.0061386
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    References listed on IDEAS

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    1. Erik D. Andrulis & Aaron M. Neiman & David C. Zappulla & Rolf Sternglanz, 1998. "Perinuclear localization of chromatin facilitates transcriptional silencing," Nature, Nature, vol. 394(6693), pages 592-595, August.
    2. Peter Fraser & Wendy Bickmore, 2007. "Nuclear organization of the genome and the potential for gene regulation," Nature, Nature, vol. 447(7143), pages 413-417, May.
    3. Yinhai Wang & Craig Ledgerwood & Claire Grills & Denise C Fitzgerald & Peter W Hamilton, 2012. "A Robust Co-Localisation Measurement Utilising Z-Stack Image Intensity Similarities for Biological Studies," PLOS ONE, Public Library of Science, vol. 7(2), pages 1-15, February.
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