PubMed Health⌕ Search

Biomedical subjects

W A Giaretti

Publications and source records attributed to W A Giaretti.

4 recordsLinked to original sources

Correlation between chromatin morphology as derived by digital image analysis and autoradiographic labeling pattern.

In order to interpret the Feulgen-dependent chromatin morphology on a functional basis, we performed model experiments in which labeling with 14C-thymidine and 14C-uridine was used as a functional parameter. Using a relocation facility, information on either DNA or RNA, labeling intensity of a cell was added to the parameters of image analysis by measuring the same cell by scanning photometry after Feulgen staining. The Feulgen-stained nuclei were interactively sampled and automatically segmented. Most of the textural information was gained from a flat texture image obtained by subtracting the original image from a median-filtered image. In addition to the autoradiographic features, visually recognizable differences in nuclear morphology, such as the number of nucleoli and the level of condensed (inactive) and diffuse (active) regions of the chromatin, were also correlated with textural parameters. Using the supervised cluster analysis method, an attempt was made to establish a correlation between visual nuclear morphology and autoradiographic labeling intensity that improved the functional understanding of the Feulgen features. Our results further clarify the supramolecular chromatin structure and its dynamics during specific transitions in the cell cycle, namely the G0-G1, G1-S and S-G2 transitions; this information may become useful in diagnostic procedures.

Animals↗

Multiparameter nuclear morphology of the G0-G1 transition.

Image analysis was performed using Feulgen-stained smears of L cells that were either confluent or had received a nutritional stimulus to proliferate. The cells were grown directly on slides in order to minimize damage to them during manipulation and to avoid trypsin treatment. A series of experiments was performed by varying the dose and the time of the stimulus. The nuclei were interactively sampled and automatically segmented. Most of the information about the fine chromatin structure was gained after subtracting the original image from a median-filtered image. The results indicate that there were changes in the Feulgen-stained nuclear morphology and densitometry shortly after transition of quiescent G0 cells to the proliferating G1 compartment. Multidimensional cluster analysis provided model-independent discrimination between the resting and the stimulated cells; the latter presented increased nuclear dispersion, indicating increased template activity of the cells. The results obtained are applicable both to obtaining a better understanding of the mechanisms of regulation during the G0-G1 transition and to deriving morphologic criteria applicable in diagnostic routines.

Animals↗

Concomitant changes in chromatin structure and proliferative activity in differentiating human erythroblasts.

During the passage of erythroblasts down the pathway of maturation, proliferative activity is increasingly slowed and nuclear chromatin increasingly condensed. The aim of the present study was to describe this chromatin condensation by means of textural features and to find correlations between the changes in nuclear structure and the proliferative activity. As a parameter of proliferative activity, the rate of DNA synthesis was investigated in individual cells by utilizing the technique of quantitative 14C-autoradiography. The nuclear fine structure was analyzed in the same cells, after removal of the silver grains and then staining by the Feulgen reaction, by scanning cytophotometry and the application of textural parameters as used in early cancer detection. Among several features, the skewness of the distribution of optical densities proved to be the best parameter for describing the overall morphologic progression during erythroblast maturation. It was almost completely independent of the actual stage of the cells in the cell cycle. The best correlation with the rate of DNA synthesis was found to be the Laplace-filtered optical densities, a parameter that can be interpreted as expressing the "graininess" of chromatin. Since changes in the template activity related to hemoglobinization are also associated with structural changes in chromatin, the relationship between chromatin structure and proliferative activity in erythroblasts is regarded as causal only to a limited extent.

Adult↗