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PubMed1998

Nuclear and nucleolar image analysis of human breast epithelial cells transformed by benzo[a]pyrene and transfected with the c-Ha-ras oncogene.

Changes in nuclear and nucleolar morphometric parameters were investigated by image analysis procedures in human breast MCF-10F epithelial cells expressing different stages of the tumourigenic progression after benzo[a]pyrene (BP) transformation (BP1, BP1-E, and BP1-E1 cell lines), and additionally transfected with the c-Ha-ras oncogene (BP1-Tras cell line). Nuclear pleomorphism was evident in all the transformed cells. The analysis of different morphometric parameters did not show a clear relationship between specific nuclear and nucleolar changes and the expression of the different stages of the tumourigenesis, with the exception of the nucleolar size, which could be associated to the expression of the tumourigenic phenotype, and a nucleolar area/nuclear area ratio, which discriminated the immortalized, the transformed, and the tumourigenic phenotypes from one another. The nuclear morphometric data established for the BP-transformed cells and for the cells additionally transfected with the c-Ha-ras oncogene were suggestive of complex and distinct morphofunctional mechanisms involving the in vitro transformation of the MCF-10F cells. The nuclear changes found in the BP1-Tras cell line were assumed to be related to the additional effects and/or enhanced genomic instability induced by transfection with the ras oncogene.

Benzo(a)pyrene↗
PubMed1992

Circulating antibodies against faecal bacteria assessed by immunomorphometry: combining quantitative immunofluorescence and image analysis.

A new technique to study the prevalence of circulating antibodies directed against different morphological groups ('morphotypes') of bacteria in fresh faeces is presented. The technique combines quantitative indirect immunofluorescence with digital image analysis. Plasma antibody titres and patterns of IgA, IgG and IgM isotype against morphotypes of faecal bacteria were determined in ten healthy individuals.

Adult↗
PubMed1994

Video image analysis of labile zinc in viable pancreatic islet cells using a specific fluorescent probe for zinc.

We used an intracellular zinc-specific fluorophore, Zinquin, in conjunction with fluorescence video image analysis, to reveal labile zinc in pancreatic islet cells, which concentrate this metal for use in synthesis, storage, and secretion of insulin. Zinquin vividly demonstrated zinc in the islet cell secretory granules, which formed a brightly labeled crescent in the cytoplasm between one side of the nucleus and the plasma membrane. Lower but still appreciable amounts of zinc were detected in the remaining cytoplasm, but there was little labeling in the nucleus. Fluorescence intensity varied among islet cells, suggesting differences in zinc content. Their average fluorescence intensity greatly surpassed that of the surrounding pancreatic acinar cells in frozen sections of pancreas and in all other types of cell studied, including lymphocytes, neutrophils, fibroblasts, and erythrocytes. Less labile zinc was detected in cells of the mouse insulinoma cell line NIT-1, regardless of whether they were maintained in long-term culture in the presence or absence of exogenous extracellular zinc. Exposure of islet or insulinoma cells to a high concentration of glucose or other secretagogue decreased the content of labile zinc. Zinquin should be a useful probe for revealing changes in zinc homeostasis in islet B-cells that may be important in their dysfunction and death during diabetes.

Animals↗
PubMed1996

Quantitative immunohistochemistry of factor VIII-related antigen in breast carcinoma: a comparison of computer-assisted image analysis with established counting methods.

Microvessel density in the area of the most intense neovascularization in invasive breast carcinoma is reported to be an independent prognostic factor. The established method of enumeration of microvessel density is to count the vessels using an ocular raster (counted microvessel density [CMVD]). The vessels were detected by staining endothelial cells using Factor VIII-related antigen. The aim of the study was to compare the CMVD results with the percentage of factor VIII-related antigen-stained area using computer-assisted image analysis. A true color red-green-blue (RGB) image analyzer based on a morphologically reduced instruction set computer processor was used to evaluate the area of stained endothelial cells. Sixty invasive breast carcinomas were included in the analysis. There was no significant correlation between the CMVD and the percentage of factor VIII-related antigen-stained area (Spearman correlation coefficient = 0.24, confidence interval = 0.02-0.46). Although high CMVD was significantly correlated with poorer recurrence free survival (P = .024), percentage of factor VIII-related antigen-stained area showed no prognostic value. Counted microvessel density and percentage of factor VIII-related antigen-stained area showed a highly significant correlation with vessel invasion (P = .0001 and P = .02, respectively). There was no correlation between CMVD and percentage of factor VIII-related antigen-stained area with other prognostic factors. In contrast to the CMVD within malignant tissue, the percentage of factor VIII-related antigen-stained area is not suitable as an indicator of prognosis in breast cancer patients.

Breast Neoplasms↗
PubMed2004

Measurement of gas holes and mechanical openness in cheese by image analysis.

A method to measure the amount of the surface area of cheese slices occupied by gas holes was developed to reflect the relative gas production among different cheeses. A digital camera mounted on a copy stand with lighting was used to make digital images of each slice of cheese. A commercial digital image analysis software program was used and an algorithm was written to measure the area of the image of the cheese slice occupied by holes. The image was cropped and scanned to determine which color channel produced the best image contrast. The MATLAB program allowed the user to eliminate mechanical openness or false holes and then to scan the image to produce a percent distribution of pixels in the image as a function of pixel intensity. The user then determined a threshold value to differentiate pixels that were in holes from those representing areas with no holes. The percentage of the total surface area occupied by holes was calculated. The coefficient of variation of the method ranged from 2.43% with gas holes of about 1% of the surface of the cheese slice to a coefficient of variation of 0.92% with gas holes of about 6.8% of the surface area of the cheese slice. Examples of applications of this method are given for Emmental, Ragusano, and Cheddar cheeses. The method can be used as a tool in research studies to correlate the amount of gas production with manufacturing conditions or as a quality control tool in cheese manufacturing.

Cheese↗
PubMed2003

A new method for assessing arteriolar diameter and hemodynamic resistance using image analysis of vessel lumen.

To characterize the nonuniform diameter response in a blood vessel after a given stimulus (e.g., arteriolar conducted response), frequent serial diameter measurements along the vessel length are required. We used an advanced image analysis algorithm (the "discrete dynamic contour") to develop a quick, reliable method for serial luminal diameter measurements along the arteriole visualized by intravital video microscopy. With the use of digitized images of the arteriole and computer graphics, the method required an operator to mark the image of the two inner edges of the arteriole at several places along the arteriolar length. The algorithm then "filled in" these marks to generate two continuous contours that "hugged" these edges. A computer routine used these contours to determine luminal diameters every 20 microm. Based on these diameters and on Poiseuille's law, the routine also estimated the hemodynamic resistance of the blood vessel. To demonstrate the usefulness of the method, we examined the character of spatial decay of KCl-induced conducted constriction along approximately 500-microm-long arteriolar segments and the KCl-induced increase in hemodynamic resistance computed for these segments. The decay was only modestly fitted by a simple exponential, and the computed increase in resistance (i.e., 5- to 70-fold) was only modestly predicted by resistance increase based on our mathematical model involving measurements at two arteriolar sites (Tyml K, Wang X, Lidington D, and Oullette Y. Am J Physiol Heart Circ Physiol 281: H1397-H1406, 2001). We conclude that our method provides quick, reliable serial diameter measurements. Because the change in hemodynamic resistance could serve as a sensitive index of conducted response, use of this index in studies of conducted response may lead to new mechanistic insights on the response.

Animals↗
PubMed2000

Improved quantitative characterization of atherosclerotic plaque composition with immunohistochemistry, confocal fluorescence microscopy, and computer-assisted image analysis.

To quantitatively characterize contributions of major constituents to the composition of a given atherosclerotic plaque, we have developed an approach employing immunohistochemistry, confocal scanning laser microscopy, and computer-assisted image analysis. The method developed permits identification of plaques that are particularly vulnerable to rupture and elucidation of the nature of the composition of a given plaque, as well as the extent of luminal encroachment. Thus, it should be useful in experimental animals and ultimately in patients in delineating compositional changes in response to potentially deleterious genetic and environmentally induced factors and to potentially therapeutic interventions designed to diminish plaque vulnerability.

Animals↗
PubMed2002

Characterization of low molecular weight alkoxylated polymers using long column SFC/MS and an image analysis based quantitation approach.

The utility of low viscosity mobile phases and long chromatographic columns for complex polymer analysis is demonstrated. We use long column supercritical fluid chromatography/mass spectrometry (SFC/MS) with electrospray ionization (ESI) to characterize a variety of complex, low molecular weight polymers. When quantitative analysis is desired, the resulting three-dimensional (time, intensity, and mass-to-charge ratio [m/z]) data are converted to images. Custom image analysis software is used to detect and integrate peaks in arbitrarily defined regions of the time-m/z map. These integrated peak volumes can be used to quantitate distinct component classes of the polymer mixtures.

Journal Article↗
PubMed1990

Electron microscopic radioautographic studies on DNA synthesis in the hepatocytes of aging mice as observed by image analysis.

Quantitative changes of DNA synthesis in the hepatocytes of aging mice were studied by electron microscopic radioautography. Ultrastructural changes of labelled and unlabelled hepatocytes were estimated quantitatively by the image analysis. The results revealed that at various ages, the area of the cytoplasm and nuclei of labelled hepatocytes were more than those of the unlabelled cells. No significant changes were observed in the nucleoli. The area of the endoplasmic reticulum and mitochondria were less in the labelled hepatocytes than in the unlabelled hepatocytes. The number of the mitochondria was less in the labelled hepatocytes than in the unlabelled hepatocytes. The cell organelles of the hepatocytes that were synthesizing DNA were not well developed, as compared to those not synthesizing DNA during the postnatal development.

Aging↗
PubMedDate not supplied

Automated image analysis, histologic malignancy grading, and survival in patients with astrocytic gliomas. Prognostic significance of nuclear count and nuclear area fraction.

A number of morphometric and densitometric parameters, obtained by automated image analysis, correlate fairly well with classic histologic grading of malignancy in astrocytic gliomas. In this study the predictive value of classic histologic grading of malignancy and of two automatically retrieved nuclear parameters are compared in 199 patients treated surgically for an astrocytic glioma. A 100% follow-up was obtained. Nuclear count and nuclear area fraction correlated fairly well with survival. In patients with 25% of the lowest values of nuclear count and nuclear area fraction 5-year survival was 40%. In patients with 25% of the highest values, less than 5% survived. In contrast, subdivision of the glial tumors into four malignancy grades gave an even better prognostic accuracy. In patients with gliomas of malignancy grade I, about 60% survived for more than 15 years; in malignancy grade IV, no patients survived for more than 5 years.

Adolescent↗
PubMed1995

Areas of dormant glial fibrillary acidic protein (GFAP) immunoreactivity in the rat brain as revealed by automated image analysis of serial coronal sections.

Coronal vibratome sections of the rat brain were immunostained for glial fibrillary acidic protein (GFAP) a major cytoskeletal protein typical for astrocytes. Using computer-assisted image analysis of whole sections, a substantial heterogeneity in regional staining intensities was pointed out. The middle layers of the neocortex, the thalamus and caudate-putamen and the tectal and tegmental part of the mesencephalon were found to show no detectable staining, while peak staining intensities were measured for the pallidum, septal triangular nucleus, hippocampus, medial geniculate nucleus and interpeduncular nucleus. In some of the negative areas neural lesioning induced the appearance of GFAP immunoreaction parallel to an up-regulation of GFAP synthesis. On this basis it was assumed that GFAP immunoreactivity is dormant in these astrocytes and plastic changes in the neuropil trigger its expression.

Animals↗
PubMed1992

[Computerized image analysis for retinal nerve fiber layer: a preliminary report].

The principle and method for analysing the white-black photographs of retinal nerve fiber layer (RNFL) by using the technique of computerized image analysis were introduced. An objective and quantitative result of RNFL on the white-black photograph was initially obtained, and using the technique of fulse colour, each of the RNFL could be showed, which seem to have more advantages than observing RNFL on a white-black photograph.

Anthropometry↗
PubMed1993

Effect of doxorubicin, daunorubicin, and idarubicin on the growth rate of human leukemia cells (K562) studied with image analysis.

The effects of doxorubicin, daunorubicin, and idarubicin on the growth of K562 cells were investigated by monitoring the formation of individual colonies in semi-solid culture employing an automated image analysis system. Drug effects were evaluated using short-term exposure (2 h) at clinically achievable drug plasma concentrations. Following drug exposure, heterogeneity in growth patterns was observed which was categorized into three distinct types; (1) continuous growth at a decreased growth rate; (2) limited growth; (3) no growth. In addition to a concentration-dependent decrease of the growth fraction, a reduction of the growth rate of the continuously growing colonies was observed. At equitoxic drug concentrations, comparable changes in growth rate were observed for each of the three anthracyclines studied. It is demonstrated that the fraction of cells which escape drug-induced growth arrest with therapeutically achievable drug concentrations display a significant decrease in growth rate.

Cell Division↗
PubMed1989

[Neuroradiological image analysis and prognostic factors of malignant gliomas in the cerebral hemisphere of adults].

54% cases of malignant gliomas followed more than 2 years were studied about prognostic factors focusing on neuroradiologic image analysis in the two groups classified survival cases over 2 years and dead cases within 2 years after surgical treatment. 21% cases of malignant gliomas having tumor infiltration or edema to contralateral hemisphere on CT and MRI before treatment were dead within 2 years. 80% of recurrent malignant gliomas had abnormal shadow on CT due to tumor recurrence in primary tumor site or ventricular wall and/or contralateral hemisphere continuing with there. Remote metastasis discontinuing with primary site was demonstrated in only 20% of them. Over 90% of glioblastoma multiforme showing irregular tumor stain with arteriovenous fistula on angiogram were dead within 2 years, whereas in the cases without tumor stain the alive cases more than 2 years increased comparing with those having tumor stain.

Adult↗
PubMed1997

Image analysis of the duodenal endocrine cells in mice with particular regard to optical densitometry.

The endocrine cells in the murine proximal duodenum have been investigated by means of immunohistochemistry and computerized image analysis. Five endocrine cell types were identified, namely secretin-, gastric inhibitory polypeptide (GIP)-, gastrin-CCK-, somatostatin- and serotonin immunoreactive cells. The number of endocrine cells/mm3 epithelial cells was estimated and the cell secretory index (CSI) for different endocrine cell types was determined. Furthermore, the optical density of the cellular immunoreactivity and the immunoreactive area in the cell were determined and an index, cell immunoreactivity content was estimated as the optical density multiplied by the immunoreactive area. It has been suggested that the use of this index might better reflect the cellular peptide/amine content than does the CSI. Serotonin-immunoreactive cells were the predominant endocrine cell type, followed by gastrin/CCK-immunoreactive cells. The numbers of secretin-, GIP- and somatostatin immunoreactive cells were almost identical. All endocrine cell types were present both in crypts and in villi, but were, more numerous in the crypts, except for secretin which was more frequent in the villi.

Animals↗
PubMed1994

Novel application of image analysis to the detection of spongiform change.

Spongiform change is the characteristic neuropathologic abnormality within the brain in Creutzfeldt-Jakob disease, the most common human spongiform encephalopathy, but traditional methods of evaluating this abnormality are subjective and laborious. In order to overcome these difficulties, a quantitative method of assessment of spongiform change was devised and implemented using image analysis techniques. Successful validation of this system was performed by subjective and objective comparison with a neuropathologist's assessment. In a test-set of 50 microscopic images a strong positive rank correlation (Rs = 0.91) was established between the image analyzer and the neuropathologist. This system should facilitate large-scale analysis of different patterns of spongiform change in cases of human spongiform encephalopathy.

Brain↗