Objective assessment of changes in nuclear morphology and cell distribution following induction of apoptosis.
Objective assessment of changes in nuclear morphology and cell distribution following induction of apoptosis.
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DOI：
10.1186/1746-1596-9-92
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发表时间：
2014-05-12
影响因子：
2.6
通讯作者：
Utheim TP
中科院分区：
文献类型：
--
作者：
Eidet JR;Pasovic L;Maria R;Jackson CJ;Utheim TP
客观测量诱导凋亡后细胞核形态和细胞分布的变化。将自发永生化的视网膜色素上皮细胞系（ARPE-19）在含有10%胎牛血清的DMEM/F12中培养三天，随后在星形孢菌素中孵育24小时以诱导凋亡。未在星形孢菌素中孵育的细胞用作对照。Caspase-3在凋亡细胞中的表达通过定量免疫荧光证实。细胞核用DAPI复染。使用ImageJ软件进行核形态和细胞分布的评估。统计学分析包括Student t检验和Pearson相关系数。最近邻分析用于评估细胞核分布。与对照组相比，星形孢菌素孵育的细胞中Caspase-3的表达增加了471% ± 182%（P = 0.014）。与对照组相比，星形孢菌素孵育培养物中的细胞具有较小的平均核面积（68% ± 5%; P < 0.001）和核周长（78 ± 3%; P < 0.001），而核形状因子较大（110% ± 1%; P < 0.001）。来自星形孢菌素组（R = 1.12 ± 0.04; P < 0.01）和对照组（R = 1.28 ± 0.03; P < 0.01）的细胞核在整个培养物中均匀分布，从而证明了非聚集和非随机的细胞分布。然而，与对照相比，星形孢菌素孵育组具有显著较低的R值（P = 0.002），这表明在诱导细胞凋亡后向细胞簇集移动。Caspase-3的表达与以下形态学指标显著相关：细胞核周长除以形状因子（r = -0.475; P < 0.001），核面积除以形状因子（r = -0.470; P < 0.001），核周长（r = -0.469; P < 0.001），核面积（r = -0.445; P < 0.001）、核形状因子（r = 0.410; P < 0.001）和核面积乘以形状因子（r = -0.377; P < 0.001）。Caspase-3阳性凋亡细胞表现出可以使用免费获得的ImageJ软件客观定量的形态学特征。一种新的形态学指标，定义为核周长除以形状因子，表现出最强的相关性与caspase-3的表达。本文的虚拟幻灯片可以在这里找到：http://www.diagnosticpathology.diagnomx.eu/vs/3271993311662947
To objectively measure changes in nuclear morphology and cell distribution following induction of apoptosis. A spontaneously immortalized retinal pigment epithelial cell line (ARPE-19) was cultured for three days in DMEM/F12 with 10% fetal bovine serum followed by 24 hours incubation in staurosporine to induce apoptosis. Cells that were not incubated in staurosporine served as control. Caspase-3 expression in apoptotic cells was demonstrated by quantitative immunofluorescence. Nuclei were counterstained with DAPI. Assessments of nuclear morphology and cell distribution were performed using ImageJ software. Statistical analyses included Student’s t-test and Pearson’s correlation coefficient. Nearest neighbor analysis was used to assess cell nuclei distribution. Caspase-3 expression in staurosporine-incubated cells increased by 471% ± 182% compared to control (P = 0.014). Relative to the control, cells in the staurosporine-incubated cultures had smaller average nuclear area (68% ± 5%; P < 0.001) and nuclear circumference (78 ± 3%; P < 0.001), while nuclear form factor was larger (110% ± 1%; P < 0.001). Cell nuclei from the staurosporine-group (R = 1.12 ± 0.04; P < 0.01) and the control (R = 1.28 ± 0.03; P < 0.01) were evenly spaced throughout the cultures, thereby demonstrating a non-clustered and non-random cell distribution. However, the staurosporine-incubated group had a significantly lower R-value compared to the control (P = 0.002), which indicated a move towards cell clustering following induction of apoptosis. Caspase-3 expression of each individual cell correlated significantly with the following morphological indicators: circumference of the nucleus divided by form factor (r = -0.475; P < 0.001), nuclear area divided by form factor (r = -0.470; P < 0.001), nuclear circumference (r = -0.469; P < 0.001), nuclear area (r = -0.445; P < 0.001), nuclear form factor (r = 0.410; P < 0.001) and the nuclear area multiplied by form factor) (r = -0.377; P < 0.001). Caspase-3 positive apoptotic cells demonstrate morphological features that can be objectively quantified using freely available ImageJ software. A novel morphological indicator, defined as the nuclear circumference divided by form factor, demonstrated the strongest correlation with caspase-3 expression. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/3271993311662947
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影响因子：
2.6
作者：
Belhomme, Philippe;Oger, Myriam;Herlin, Paulette
通讯作者：
Herlin, Paulette
影响因子：
1.6
作者：
TARNOWSKI, BI;SPINALE, FG;NICHOLSON, JH
通讯作者：
NICHOLSON, JH
DOI：
10.1016/j.brainresprot.2004.04.001
发表时间：
2004-08-01
期刊：
BRAIN RESEARCH PROTOCOLS
影响因子：
--
作者：
Daniel, B;DeCoster, MA
通讯作者：
DeCoster, MA
影响因子：
2.6
作者：
Kayser K;Schultz H;Goldmann T;Görtler J;Kayser G;Vollmer E
通讯作者：
Vollmer E
迈向自动化的虚拟幻灯片筛选：在互联网中实施的基于自动化组织的虚拟诊断的理论考虑和实际经验。
影响因子：
2.6
作者：
Kayser, Klaus;Radziszowski, Dominik;Bzdyl, Piotr;Sommer, Rainer;Kayser, Gian
通讯作者：
Kayser, Gian