Premium
Characterizing the three‐dimensional organization of telomeres
Author(s) -
Vermolen B. J.,
Garini Y.,
Mai S.,
Mougey V.,
Fest T.,
Chuang T. C.Y.,
Chuang A. Y.C.,
Wark L.,
Young I. T.
Publication year - 2005
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20159
Subject(s) - telomere , computational biology , biology , evolutionary biology , genetics , dna
Background Quantitative analysis can be used in combination with fluorescence microscopy. Although the human eye is able to obtain good qualitative results, when analyzing the spatial organization of telomeres in interphase nuclei, there is a need for quantitative results based on image analysis. Methods We developed a tool for analyzing three‐dimensional images of telomeres stained by fluorescence in situ hybridization in interphase nuclei with DNA counterstained with 4′,6‐diamidino‐2‐phenylindole. After deconvolution of the image, we segmented individual telomeres. From the location of the telomeres we derived a distribution parameter ρ T , which indicated whether the telomeres were in a disk (ρ T ≫ 1) or not (ρ T ≈ 1). We sorted mouse lymphocyte nuclei and measured ρ T . We also performed a bromodeoxyuridine synchronous cell sorting experiment on live cells and measured ρ T at several instances. Results Measuring ρ T for nuclei in G0/G1, S, and G2 produced 1.4 ± 0.1, 1.5 ± 0.2, and 14 ± 2, respectively, showing a significant difference between G2 and G0/G1 or S. For the bromodeoxyuridine synchronous cell sorting experiment, we found a cell cycle dependency of ρ T and a correlation between ρ T and an observer. Conclusions In this study we present a quantitative method to characterize the organization of telomeres using three‐dimensional imaging, image processing, and image analysis. © 2005 International Society for Analytical Cytology