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Characterization of chromatin domains by 3D fluorescence microscopy: An automated methodology for quantitative analysis and nuclei screening
Author(s) -
Cantaloube Sylvain,
Romeo Kelly,
Le Baccon Patricia,
Almouzni Geneviève,
Quivy JeanPierre
Publication year - 2012
Publication title -
bioessays
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.175
H-Index - 184
eISSN - 1521-1878
pISSN - 0265-9247
DOI - 10.1002/bies.201100188
Subject(s) - chromatin , heterochromatin , computational biology , characterization (materials science) , fluorescence microscope , fluorescence , microscopy , biological system , biology , resolution (logic) , computer science , genetics , nanotechnology , artificial intelligence , physics , dna , materials science , optics
Fluorescence microscopy has provided a route to qualitatively analyze features of nuclear structures and chromatin domains with increasing resolution. However, it is becoming increasingly important to develop tools for quantitative analysis. Here, we present an automated method to quantitatively determine the enrichment of several endogenous factors, immunostained in pericentric heterochromatin domains in mouse cells. We show that this method permits an unbiased characterization of changes in the enrichment of several factors with statistical significance from a large number of nuclei. Furthermore, the nuclei can be sorted according to the enrichment value of these factors. This method should prove useful to monitor events related to changes in the amount, rather than the presence or absence, of any factor. By adapting a few parameters, it could be extended to other nuclear structures and the benefit of using available software will permit its use in many biological labs.