Nuclear shape defects are a distinguishing characteristic in laminopathies, cancers, and other pathologies. Correlating these defects to the symptoms, mechanisms, and progression of disease requires unbiased, quantitative, and high-throughput means of quantifying nuclear morphology. To accomplish this, we developed a method of automatically segmenting fluorescently stained nuclei in 2D microscopy images and then classifying them as normal or dysmorphic based on three geometric features of the nucleus using a package of Matlab codes. As a test case, cultured skin-fibroblast nuclei of individuals possessing LMNA splice-site mutation (c.357-2A>G), LMNA nonsense mutation (c.736 C>T, pQ246X) in exon 4, LMNA missense mutation (c.1003C>T, pR335W) in exon 6, Hutchinson-Gilford Progeria Syndrome, and no LMNA mutations were analyzed. For each cell type, the percentage of dysmorphic nuclei, and other morphological features such as average nuclear area and average eccentricity were obtained. Compared to blind observers, our procedure implemented in Matlab codes possessed similar accuracy to manual counting of dysmorphic nuclei while being significantly more consistent. The automatic quantification of nuclear defects revealed a correlation between in vitro results and age of patients for initial symptom onset. Our results demonstrate the method's utility in experimental studies of diseases affecting nuclear shape through automated, unbiased, and accurate identification of dysmorphic nuclei.
Fig3_All_and_Defective_Statistics
Sigma-Plot spreadsheet with the data for Figure 3 of the manuscript. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
Fig3_All_and_Defective_Statistics_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for Figure 3 of the manuscript. )
Fig4a_Data
Sigma-Plot file with data for figure 4a of the manuscript. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
Fig4a_Data_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for figure 4a of the manuscript.)
Fig4b_Data
Sigma-plot data file for figure 4b in the manuscript. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
Fig4b_data_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for figure 4b in the manuscript.)
FigS5
Sigma-plot file with data for Fig. S5. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
FigS5_Export
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for Fig. S5. )
Fig3_Graphs
Sigma plot file containing data for other all families shown in Fig. 3. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
JasonPaper_Fig3_RedoGraphs.JNB
Figure3-All_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for all families shown in Fig. 3. )
JasonPaper_Fig3_RedoGraphs_Excel.xlsx
LaminAC Statistics
Sigma Plot Data file containing the data for comparing lamin and dapi stains. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
LaminAC Statistics_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for comparing lamin and dapi stains. )
Validation Stats
Sigma-Plot file containing data for validation statistics in Table 1. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
Validation Stats_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for validation statistics in Table 1.)
3D-PLOT-DATA-Fig2
Sigma-plot file containing data for Fig. 2 3D plots. An excel file containing just the data-sheets is also provided under the same name for those with no access to jnb files.
3D-PLOT-DATA.JNB
3D-PLOT-DATA_Fig2_Excel
An excel file containing just the data-sheets provided under the same name for those with no access to jnb files. (Data for Fig. 2 3D plots.)
3D-PLOT-DATA_Excel.xlsx