Nucleus image properties assessed by video image analysis in mouse hepatocytes under a short lysis for extended chromatin fiber formation

Vidal, Benedicto de Campos; Moraes, Alberto S.; Mello, Maria Luiza S.

doi:10.1002/cyto.a.20339

Cited by 7 publications

(6 citation statements)

References 28 publications

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“…Two threshold values determined which gray value range of the image Input is retained or deleted in the image Output. Threshold values (gray values) named Low (L) and High (H) were determined by moving the borders in the gray value histogram such that nuclear images appeared well segmented from each other and from the background (18). Since 'green' was selected as the parameter 'color', the pixels inside the gray value range (L, H) were displayed in this color.…”

Section: Cells and Treatmentsmentioning

confidence: 99%

“…The software provided quantitative information on 1) geometric parameters such as nuclear total area (μm 2 ), nuclear absorbing area (μm 2 ), nuclear perimeter (μm), and nuclear feret ratio (minimal feret/maximal feret, as an indication of the elliptical shape of the nuclear absorbing image), 2) densitometric parameters such as mean gray value per nucleus [subsequently converted into absorbances or optical density (OD)], and integrated optical density (IOD), and 3) textural parameters, such as standard deviation of the total densitometric values per nucleus or absorbance variability per nucleus (SDtd), and nuclear entropy (E). SDtd reflects the variability in the degree of chromatin packing state per Feulgen-stained image (18,19). Entropy is considered the number of bits necessary to store the densitometric values per nucleus image or amount of gray value variability in a nucleus.…”

Section: Cells and Treatmentsmentioning

confidence: 99%

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17-β-estradiol affects nuclear image properties in MCF-10F human breast epithelial cells with tumorigenesis

Mello¹,

Russo²,

Russo³

et al. 2007

Oncol Rep

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Treatment of the human breast epithelial cells MCF-10F with 17-ß-estradiol (E2) induces transformation and tumorigenesis. E2-transformed MCF-10F cells are known to exhibit progressive loss of ductulogenesis, and invasive and tumorigenic phenotypes. Although DNA amounts and chromatin supraorganization change in E2-transformed MCF cells, no comparative study has yet been undertaken in the resulting cells selected for aggressive invasiveness (C5) and tumor generation in a heterologous host. The aim of this study was thus to determine whether changes in Feulgen-DNA content and chromatin supraorganization are involved during E2-induced transformation and tumorigenesis of the MCF-10F cells. Image analysis was performed for nontransformed and E2-transformed MCF cells, highly invasive cells (C5), and for cell lines (C5-A6-T6 and C5-A8-T8) derived from tumors generated by injection of C5 cells in SCID mice. A decrease in Feulgen-DNA amounts and nuclear sizes induced by E2 treatment was accented with selection of the highly invasive tumorigenesis potential. However, in the tumor-derived cells a high variability in cellular phenotypes resulted inclusively in near-polyploidy. Significant changes in textural parameters, including nuclear entropy, indicated chromatin structural remodeling with advancing tumorigenesis. An increased variability in the degree of chromatin packing states in the E2-transformed MCF cells is followed by reduction in chromatin condensation and in contrast between condensed and noncondensed chromatin in the highly invasive C5 cells and tumor-derived cell lines. Studies on epigenetic mechanisms involving DNA methylation and/or the histone code would contribute to a better interpretation of the chromatin supraorganization changes reported herein.

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Section: Cells and Treatmentsmentioning

confidence: 99%

Section: Cells and Treatmentsmentioning

confidence: 99%

17-β-estradiol affects nuclear image properties in MCF-10F human breast epithelial cells with tumorigenesis

Mello¹,

Russo²,

Russo³

et al. 2007

Oncol Rep

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show abstract

“…Among the various image analysis textural parameters estimated in Feulgen-stained cells, inclusive in transformed MCF-10F cells, entropy has been shown to be relevant for discrimination of changes in chromatin supraorganization (2,(5)(6)(7). Nuclear entropy as studied by video image analysis has been defined as the number of bits necessary to store the densitometric values per nucleus image or the amount of gray value variability contained in a nucleus (5,7,9).…”

Section: Introductionmentioning

confidence: 99%

“…Nuclear entropy as studied by video image analysis has been defined as the number of bits necessary to store the densitometric values per nucleus image or the amount of gray value variability contained in a nucleus (5,7,9). This parameter measures the complexity of an image (10), such that a high entropy value for Feulgen-stained cells is found when there is a high contrast between condensed and non-condensed chromatin, whereas a low entropy value represents a homogeneously packed chromatin state (fairly constant gray levels) (7,8) (Fig.…”

Section: Introductionmentioning

confidence: 99%

Entropy of Feulgen-stained 17-β-estradiol-transformed human breast epithelial cells as assessed by restriction enzymes and image analysis

Mello

Russo²,

Russo³

et al. 2009

Oncol Rep

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Abstract. MCF-10F human breast epithelial cells when transformed with 17-ß-estradiol (E2) give rise to highly invasive C5 cells that generate adenocarcinomas in SCID mice. From these tumors, cell lines such as C5-A6-T6 and C5-A8-T8 have been derived. Variable patterns of chromatin supraorganization have been demonstrated for these cells during the transformation/tumorigenesis progress, when assessing chromatin entropy by image analysis in Feulgenstained preparations. Since epigenetic dysregulation might contribute to the chromatin textural repatterning in transformed MCF-10F cells, the association of the variable chromatin packing states with global DNA methylation was investigated in these cells after their treatment with restriction enzymes followed by Feulgen staining and chromatin entropy evaluation by image analysis. The results indicate that although -C m CGG-sequences may affect chromatin supraorganization in some of the analyzed cell types (perhaps due to localized hypermethylation), not all the chromatin condensation patterns in these cells with transformation and/ or tumorigenesis are associated with DNA methylation (e.g. E2 cells). Chromatin supraorganization remodeling in C5-A6-T6 and C5-A8-T8 cells may be attained by different mechanisms, with C5-A6-T6 chromatin packing states perhaps being associated with local DNA hypermethylation or other epigenetic factors, and C5-A8-T8 likely being associated with global DNA hypomethylation, as reported in the literature for other cell types. Thus, we assume that a variable epigenetic modulation affecting the higher-order packing states of chromatin in the estrogentransformed MCF-10F cell model could be evident with the chromatin entropy study by image analysis.

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“…Image‐based analysis of nuclear morphometry is a key problem due to the important roles that the cell nucleus plays in biology. Nuclear morphology, and associated changes, have been studied extensively lately (18–20). In this issue Rohde et al (p 341) discussed the difficulties associated with the analysis of nuclear shape since the shape spaces are nonlinear.…”

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confidence: 99%

New trends in cytometry in the era of systems biology

2008

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Nucleus image properties assessed by video image analysis in mouse hepatocytes under a short lysis for extended chromatin fiber formation

Cited by 7 publications

References 28 publications

17-β-estradiol affects nuclear image properties in MCF-10F human breast epithelial cells with tumorigenesis

17-β-estradiol affects nuclear image properties in MCF-10F human breast epithelial cells with tumorigenesis

Entropy of Feulgen-stained 17-β-estradiol-transformed human breast epithelial cells as assessed by restriction enzymes and image analysis

New trends in cytometry in the era of systems biology

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