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PubMed1991

DNA analysis of cardiac myxomas: flow cytometry and image analysis.

Cardiac myxoma is the most common primary tumor of heart, but there is a longstanding controversy over whether it is a true neoplasm or a reactive lesion. We analyzed 24 cardiac myxomas from 22 patients: 22 by DNA flow cytometry and five by image analysis. Two myxomas were aneuploid; one of those analyzed by flow cytometry, and the other by image analysis. Proliferative fractions (S + G2/M) were high in three tumors from patients with multiple myxomas (mean, 15.9%; SD, 4.0%) as compared with 12 solitary uncomplicated myxomas (mean, 7.7%; SD, 6.0%). S-phase and proliferative fractions were low in embolic, recurrent, and solitary myxomas. The presence of aneuploidy in some myxomas supports a neoplastic origin for this tumor.

Adult↗
PubMed1993

Measurement of osteoclasts and bone resorption by automated image analysis.

An automated image analysis method is described for measurement of osteoclasts and resorbing surface in calcified bone. Osteoclasts, osteoid, and mineralized bone were measured in a single section, reacted for acid phosphatase activity, and then stained with orange G and light green stain. Three images were acquired of each field with a monochromatic camera at illuminating wavelengths of 635, 540, and 480 nm (selected using a stage monochromator). These wavelengths were chosen according to the absorption spectra of the different image components to maximize absorption differences between osteoclasts (red), mineralized bone (blue/green), and osteoid (orange). These components were then discriminated according to operator-defined ranges of color density (mineralized bone) or color fraction (osteoid and osteoclasts). A gray level coded segmented image was produced, from which was determined the area, perimeter, and number of each component and the length of contact zones with the marrow and between these components. The method was evaluated by twice measuring 10 bone sections from patients with end-stage liver failure awaiting liver transplantation. The method was quite reproducible, with coefficients of variation varying between 4% for bone volume (% tissue volume) and 22% for osteoid surface (% bone surface). The sections were also measured using a previously established semiautomated method. Coefficients of variation between methods were higher varying between 4% for bone volume (% tissue volume) and 56% for osteoid volume (% bone volume). The automated method gave a substantial time saving compared to the semiautomated method. An interactive technique was used in adjacent sections to evaluate tetracycline labeling and osteoblast surfaces.

Adolescent↗
PubMed1983

Differentiation of human T and B peripheral blood lymphocytes by high-resolution cell image analysis.

Automated cell image analysis of light and electron microscopic pictures was used for differentiation of nonlabeled lymphocytes in blood smears and in smears of purified lymphocyte suspensions. The percentages of T and B lymphocytes were determined by a two-step rosette assay with sheep red blood cells (T cells) and an immunofluorescence assay with FITC-labeled antihuman globulin (B cells). Images from 1,400 Feulgen-stained and 12,000 Pappenheim-stained cells were analyzed. Various classification methods allowed two lymphocyte subpopulations to be discriminated at the light and electron microscopic levels on the basis of different visual and subvisual morphologic features. As found by immunologic methods, morphologically determined subpopulations corresponded to T and non-T cells, with no further differentiation of non-T cells into B or null cells possible. The results allow the conclusion that there are morphologic differences between human T and non-T cells, with the differences distinguishable from individual variations as well as from alterations induced by sample preparation.

B-Lymphocytes↗
PubMed1983

Histochemical labeling of drug receptors in schistosomes: quantitation using computer image analysis techniques.

Computer image analysis techniques were used to quantitate the histochemical labeling of schistosomes with dimethylaminonaphthalene-5-sulfonaminoethyl methylamine hydrochloride (DDNS), a fluorescent analog of acetylcholine (ACh). Dose-response curves, consistent with previous experiments, were obtained for DDNS, and for blockage of DDNS labeling, by trifluoperazine and mecamylamine.

Animals↗
PubMed1998

Physiological investigations by image analysis.

Modern quantitative image analysis has been extensively used to characterize the morphology of microorganisms, especially those of the filamentous type. More recently physiological features have been quantified, making use of classical stains as well as fluorescent dyes. The potential of the technique is illustrated by a detailed analysis of the differentiation of Streptomyces ambofaciens. Three different staining procedures have been used to monitor the thinning and septation of hyphae (with propidium iodide staining), the leakage of cellular components through the membrane (with carbol gentian violet staining) and the respiration (with INT staining) in a batch submerged culture.

Gentian Violet↗
PubMed1995

Comparative morphometric studies of benign and malignant intraductal proliferative lesions of the breast by computerized image analysis.

Using computerized image analysis, we newly devised a method for automatic quantitative evaluation of nuclear arrangements in variable proliferative intraductal (cribriform) lesions of the breast, by calculating angles of longest nuclear diameter to a horizon (LNDA). Internuclear correlation in the cribriform lesions was thus examined, comparing 22 benign and 62 malignant cases. In the malignant lesions, nuclear arrangements tended to be multidirectional, probably because of vertical nuclear arrangements toward acinar lumens as reflected by the wide distribution of LNDAs. Conversely, in the benign lesions, groups of nuclei tended to arrange toward one certain direction, forming a complex streaming pattern with the distribution of LNDAs usually producing a distinctive peak. Other conventional nuclear features, such as nuclear area, nuclear perimeter, shortest nuclear diameter (SND), and leading variables were also quantitatively assessed for multivariate analysis. It was noted that nuclear arrangements could represent a favorable discriminator. Linear discriminant function could classify 66 of the total 84 cases (78.6%) as being consistent with the diagnosis of pathologists. We conclude that internuclear arrangement could be a useful discriminating variable in benign and malignant breast lesions for the further development of an automatic analytic system.

Adenocarcinoma↗
PubMed1993

Erosion by soft drinks of rat molar teeth assessed by digital image analysis.

A digital imaging analysis (DIA) procedure for recording and assessing erosive changes on teeth was developed and evaluated against an older subjective method. Results after exposure of rats' teeth to various soft drinks showed that the DIA system was superior to the conventional method, with improved precision. The system provided a direct and highly sensitive means of quantifying the erosion of dental hard tissues by three different soft drinks, showing that it could be used to evaluate erosiveness and other dental damage. After 6 weeks' exposure to soft drinks in the diet, the area of intact enamel on the lingual surface of the first mandibular molar averaged 47% on apple juice, 27% on still orange and 6% on carbonated orange. The greatest exposure of dentine (36%) was on the still orange drink. Eroded enamel accounted for 72.5% of the tooth surface on the carbonated drink.

Animals↗
PubMed1995

Quantitation of fibroblast population growth rate in situ using computerized image analysis.

The development of computer-assisted image analysis has provided the technology to rapidly determine the population size of cultured cell monolayers in situ. We have adapted this technology to determine the population growth rate of cultured fibroblasts for use in a high-replicate format. Human lung fibroblasts were seeded into 1/2 A 96-well plates that had one-half the culture area of standard 96-well plates. The cells were cultured in medium supplemented with different concentrations of FBS and on days 0, 1, 2, 3, 5, and 7, and their nuclei were stained with propidium iodide. A microscopic field representing one-quarter of a well of fluorescent nuclear images was captured onto a Macintosh computer, and the number of nuclei were counted using an image analysis software program. There were no significant differences between the number of nuclei counted manually and the number counted using computer-assisted software, until day 7 where the cells were multilayered (P < 0.05). This image analysis method was compared to other assays typically used to estimate cell proliferation or population size, namely hemocytometer counting, a rapid colorimetric staining assay using naphthol blue-black, and [3H]-thymidine incorporation. The growth rates derived using image analysis were in close agreement with results derived from hemocytometer counts and [3H]-thymidine incorporation. However, the growth rates of cells grown in high concentrations of FBS as determined using naphthol blue-black were substantially lower than results from image analysis. We conclude that this adaptation of computer-assisted image analysis provides a method to derive accurate growth curves by directly counting the number of cells in a large number of replicates.

Cell Count↗
PubMed1995

Nuclear characterization and G2M ploidy in human brain tumors using semiautomated image analysis.

We used image analysis to study nuclear morphometry and DNA content in relation to time to tumor progression in a series of 88 patients with brain tumors. Clinical follow-up was obtained for 73 patients. The patients with diploid tumors had a longer time to tumor progression than those with triploid, tetraploid, or hypertetraploid tumors. Mean SG2M-DNA indices (DIs) increased significantly with a increase in mean DIs in all tumors. The mean DIs appeared to be dependent on the number of SG2M phase cells. We conclude that tumors with hypertetraploid in G2M ploidy are highly malignant. Those tumor cells have a large nuclear size, much deformity in nuclear shape, and great proliferative potential. The G2M-tetraploid tumors showed a shorter time to tumor progression when the number of SG2M fractions was large. In contrast, the G2M-hypotetraploid tumors showed a longer time to tumor progression in comparison with other tetraploid and hypertetraploid tumors, but the difference was not significant.

Aneuploidy↗
PubMed1996

Quantification of tumour-induced angiogenesis by image analysis.

Quantitative techniques for in vivo and in vitro angiogenesis were developed using an image analyser. In the in vivo study, a Millipore chamber filled with mouse sarcoma 180 (S180) cells was transplanted subcutaneously to the dorsal side of a mouse, and the area of neovascularization induced by the tumour cells was quantified by image analysis. Images of vascular networks with poor contrast had their contrast improved by Laplacean transformation. The area of vascular network was 16.9 mm2 in the control group without tumour cells and 44.2 mm2 in the group with tumour cells, demonstrating a significant increase in neovascularized area by tumour cells. In the in vitro study, migration of vascular endothelial cells was induced with conditioned media of S180 cells. Image analysis was used to count automatically the nuclei of migrated endothelial cells, which were stained violet with Giemsa's solution. This automated measurement by image analyser is expected to save labour and time. Checkerboard analysis revealed that the endothelial cell migration induced by S180-conditioned medium was due to chemotaxis. The quantitation method using an automated image analyser is valuable in evaluating the induction of neovascularization by tumours and the effect of pharmacological agents on tumour angiogenesis in vivo and in vitro.

Animals↗
PubMed1982

Early effects of chemical carcinogens as compared to induced cell proliferation. II. Automated image analysis.

Static automated image analysis was applied to study early variations of chromatin structure in Feulgen-stained liver nuclei from rats injected i.p. with a single dose of dimethylnitrosamine (DMNA), a well known hepatocarcinogen. An increase of nuclear area and a correspondent decrease of average optical density (integrated optical density/area) was observed, as compared with controls, in nuclei from rats treated with 5.4 mg/kg of DMNA. These findings, which were comparable with those induced by partial hepatectomy, indicate the existence in DMNA-treated cells of a chromatin DNA relaxation similar to the G0-G1 transition previously described for human diploid fibroblasts stimulated to proliferate. Because similar results were independently obtained by flow microfluorimetry, it seems reasonable to hypothesize that chromatin decondensation could be a prerequisite for cancer induction.

Animals↗
PubMed1996

Applications of image analysis in cell technology.

Image analysis is now a well established complement to optical microscopy, allowing routine quantification of microscopic observations. Recent method developments include location and enumeration of bacteria in solid foods, in situ microscopy and image analysis for on-line monitoring of yeast fermentations, and texture analysis of fungal colonies for subsequent transfer. Notable recent applications include studies on the pulsatile growth of hyphal apices, biochemical differentiation of fungal colonies, and simple structural differentiation of mycelia from submerged fungal cultures.

Journal Article↗
PubMed1994

Textural differences between AA and SS blood specimens as detected by image analysis.

High-resolution image analysis was employed in the analysis of round (discoid) erythrocytes from hematologically normal (AA) individuals, AA individuals with nonspecific anemia, individuals with sickle cell trait (AS), individuals with SC disease (SC), and individuals with sickle cell anemia (SS). The shape feature Form Factor (4 pi Area/Perimeter2) was used to select round cells and to exclude sickle and other abnormal cells. Textural features extracted from round cells of SS and SC patients were found to differ from those derived from cells of normal andanemic AA individuals. Two textural features, Standard Deviation of Run Length Matrix Counts and Rotation Moment of the Cooccurrence Matrix, discriminated between patients mean values from AA samples and those from SS samples. The ability of textural features to separate round cells into classes based on genotype suggests that high resolution image analysis may be an effective tool in the study and monitoring of sickle cell disease.

Adolescent↗
PubMed1983

Detecting infection-related changes in peripheral blood smears with image analysis techniques.

High-resolution image analysis has the potential to flag subtle changes in white blood cell morphology that may indicate the presence of certain diseases. A study was made of the feasibility of identifying patients with hematologic bacterial infections (sepsis) using measurements on Wright-Giemsa-stained peripheral blood smears. Neutrophils and lymphocytes from a group of patients with sepsis and from a control group were digitized, and parameters quantifying geometry, color, texture and shape were extracted. While color parameters differed the most between the infected and control samples, substantial differences in geometric, texture and shape parameters also were observed. Analysis of the data showed that individual neutrophils and lymphocytes from patients with sepsis were distinguishable from those of the control group with better than 84% accuracy. When average parameters were calculated from all cells of one type for each specimen, 100% accurate classification was obtained. These studies demonstrate that the image-analysis techniques used are sensitive enough to detect disease-related changes in cell morphology that are generally too subtle for reliable detection by the human eye. Future experiments will determine the specificity of this test for bacterial infections and will explore the possibility of using image analysis techniques on peripheral blood to detect and monitor a wide variety of diseases.

Cytodiagnosis↗
PubMed1992

[Progress in microscopic image analysis and the trend of image cytometry].

Image cytometry has recently developed prominently based on the digital imaging of cell morphology brought about by great advances in microcomputer hardware and software, electronics etc. This method is characterized by the quantitation of both the amount of intracellular bioactive materials and cell morphology, and is expected to further advance many cell analysis techniques involving automated cytology, chromosome image analysis, automated histopathology, viable cell analysis, 3D image construction, etc. The present use is still minimal, and advances in both hardware and software for cyto-histologic recognition are essentially necessary. Perhaps by the end of this century, more sophisticated and faster machines for automated cyto-histologic analysis will appear, replacing many microscopic techniques of presently subjective observation in the biomedical field.

Cell Cycle↗
PubMed1995

Cervical cell recognition and morphometric grading by image analysis.

Cervical cell recognition by morphometric image analysis was compared to human visual cell recognition on the same 6,375 cells from 40 dysplastic, CIS, invasive, and 10 normal pap smears. The experimental approach defined receiver operating characteristic (ROC) curves for morphometric image analysis which could be rigorously compared to previously established human visual cell recognition ROCs on the same cells. Overall performance was measured Az, the area under the ROC curves in the two instances. For morphometric image analysis cell recognition, Az = 0.91, and for human visual cell recognition, Az = 0.87. These results clearly demonstrated that morphometric image analysis is equivalent to experienced human observers in ability to recognize isolated cells from cervical smears. An approach was also developed to link the ROC analytic methods of this study to a cytopathological or histopathological grading system, or "scale", that could be expressed in terms of normal deviate units of morphometric descriptors. This approach has the advantage of describing the grading scale in terms of its ROC characteristics; in essence, it describes performance for that grading scale at any decision point along the scale, if used for two-category classification. Additionally, This concept provides for a uniform final scale, regardless of which cells or tissues are graded. Also, this type of grading scale would automatically adjust itself for measurement variance for different types of cells or tissue, by reference to normal cells or tissues, so that a standard reference could be maintained.

Carcinoma in Situ↗
PubMed1998

Morphometric analysis of pancreatic carcinoma by computer-assisted image analysis.

This report describes the results of applying the interactive image analysis system for the measurement of some cytological parameters corresponding to features of adenocarcinoma of the pancreas. The present experiments were carried out by means of the digital cell image analysis of haematoxilyn and eosin stained archival standard glass slides of cancer bearing and healthy patients. Four different parameters describing the morphology of nuclei and nucleoli were selected to quantitate the differences between control and malignant tissues: area, perimeter, elongation, and extension. The parameters that showed the greatest differences between cancerous and normal pancreas were: area and elongation in the case of nuclei as well as area and perimeter for nucleoli. However, the results of this study suggest that none of the four analysed parameters can be selected alone to discriminate neoplastic from normal cells, but could be used all together in diagnosis of pancreatic cancer.

Adenocarcinoma↗