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PubMed2000

Evaluation of the efficacy of potential antineoplastic drugs on tumour metastasis by a computer-assisted image analysis.

Computerised image analysis, performed on histological sections of (C57BL6/N) mouse lungs that had been intravenously (i.v.) injected with B16-F10 melanoma cells was used to develop a novel method to quantify the efficacy of potential antineoplastic drugs. This procedure allowed the evaluation of the rate of inhibition of growth and the anti-invasive capability of new molecules, thus resulting in more accurate data than that obtained from common macroscopical counting of surface metastatic foci. Several morphological parameters can be measured by this method: the percentage of tissue area occupied by metastases, which accounts for tumour implantation into the organ; the growth index, related to the size of the metastases, and the invasion index, related to the frequency of foci. These morphometric data were found to be correlated to the levels of lung hydroxyproline and transglutaminase activity, well known markers of tumour invasion and cell differentiation, respectively. The main objective of this computerised procedure was to evaluate how the tumour cell is affected in the host by the drug under investigation. The use of the method is exemplified by an analysis of the antitumour activity of some methylxanthines.

Animals↗
PubMed1977

Quantitative nuclear image analysis: differentiation between normal, hyperplastic, and malignant appearing uterine glands in a paraffin section. II. Computer assisted recognition by discriminant analysis.

Quantitative image analysis was applied to data from microscope photometry of nuclei in a paraffin section. The data were essentially the same as described in a previous publication (Baak and Diegenbach, 1977). The results from measurements on the nuclei of morphologically normal, atypical hyperplastic and carcinomatous uterine glands were used in discriminant analysis. With this method it is possible to discriminate between the three groups of nuclei. Depending on the (sub)set of the variables used, 60-70% of all nuclei are correctly classified in one of the three groups. Discrimination of one of the groups against the other two results in up to 81% correct classifications. Therefore, discriminant analysis offers a possibility of assisting diagnosis in an objective way.

Cell Nucleus↗
PubMed2003

Data reproducibility in fluorescence image analysis.

Fluorescence image analysis provides quantitative data on fluorescence in situ hybridization signals (FISH), immunofluorescence labelings, Green Fluorescent Protein (GFP) expression and microarrays. It is a valuable tool for decision making in the fields of biology and medicine. The aim of this study was to evaluate the reproducibility of fluorescence intensity measurements and standardization when acquisitions are performed under various but well defined conditions. Fluorescent intensity of standard beads (Inspeck series, Molecular Probes) was repeatedly measured using an image analyzer and automated procedures. Images were acquired using several integration times and neutral filter sets. A standardization procedure was used for expressing the data in a same unit: data were multiplied by the light attenuation factor and were divided by the CCD integration times. Results show that 1) standardization is possible 2) accurate and reliable fluorescence measurements can be obtained and 3) specimens showing large differences in fluorescence intensity can be objectively compared. Moreover fluorescent test slides including fluorochrome solutions and altuglas slides were tested for shading correction and as overall test systems.

Calibration↗
PubMed1990

A quantitative assessment of F-actin content and distribution in untreated and butyric acid treated murine melanoma B16a tumour cells: a fluorescence image analysis study.

Image analysis of phallacidin, a fluorescent stoichiometric probe to F-actin, permitted the cytoskeletal-associated actin 'F-actin' to be visualized morphologically and to be divided into two groups, diffuse and filamentous. The filamentous actin group was categorized further into four subgroups according to the intensity of the phallacidin probe. F-actin groups and subgroups of untreated cells and cells treated with 1.5 mM butyrate acid were analysed independently. Butyric acid treatment significantly increased total actin, defined as diffuse actin, plus filamentous subgroup actins 1-4. Specifically, butyric acid-treatment increased filamentous subgroup actin 1.

Actins↗
PubMed1985

Measurement of fluorescein angiograms of the optic disc and retina using computerized image analysis.

Computerized image analysis was used to quantify objectively fluorescein angiograms of the optic disc, peripapillary choroid, and retina. Techniques were developed to measure fluorescein filling rates of the optic disc and the retinal vessels and the area of fluorescein filling defects within the optic disc. Two subjects, one with glaucoma and the other with ocular hypertension, showed increases of areas of fluorescein filling defects of the optic disc on follow-up and are presented here as examples of the application of these techniques. This methodology can be applied to the longitudinal follow-up of individual patients with glaucoma and retinal diseases, as well as to cross-sectional studies of patient populations.

Adolescent↗
PubMed1987

A comparative study of Spitz nevus and nodular malignant melanoma using image analysis cytometry.

Image analysis cytometry can be used to estimate both nuclear DNA content and area in tissue sections. Since nodular malignant melanoma and Spitz nevus can show a remarkably similar light microscopic appearance, but may differ in behavior, we studied typical examples of these neoplasms to determine whether cytometric differences existed. Analysis of relative DNA content alone could not discriminate between these 2 entities in the 13 cases that we examined. However, Spitz nevi and nodular melanoma clearly differed in terms of maturation, which we defined as the difference between the mean nuclear size or mean nuclear DNA content of the uppermost and deepest melanocytes in each lesion. Maturation as defined by a decrease in mean nuclear DNA content proved highly significant (p less than 0.005). Only Spitz nevi showed a lesser DNA content in the deepest dermal cells as compared with upper dermal cells, suggesting that some Spitz nevi have an admixture of diploid and hyperdiploid cells in their upper portions, but mostly diploid cells in their deep portions. Only nodular melanoma showed higher mean DNA content in deep dermal cells as opposed to superficial dermal cells, suggesting that some nodular melanomas may either have clones of cells in their deep portions that have higher levels of ploidy, or more cells in the deep portion of melanomas may be in active phases of the cell cycle. Our study suggests that important cytometric differences exist between Spitz nevi and nodular melanoma, and that these could be exploited to develop cytometry into an adjunctive clinical technique.

Cell Nucleus↗
PubMed2005

Image analysis and in vivo imaging as tools for investigation of productivity dynamics in anthocyanin-producing cell cultures of Daucus carota.

An anthocyanin-producing suspension culture of Daucus carota (L.) cv. Flakkese was used as model system to study secondary metabolite production in cell culture at the individual cell level. An approach was set up in which growth and production of anthocyanins were investigated using a combination of biochemical analysis, image (colour) analysis and in vivo imaging. This novel approach was used to segment the culture in different subpopulations and dissect the productive process in the cell culture grown under two different conditions, known to differ mainly for oxygen supply and mixing intensity (volume of 50 ml or 20 ml in 250 ml flasks). The 20 ml batch cultures gave a higher content and yield of anthocyanins, which depended on a complex balance between events that positively or negatively affected anthocyanin production. A model is proposed in which the different ability of cells to respond to environmental stimuli and stress depends on the different amount of anthocyanins accumulated within cells.

Anthocyanins↗
PubMed1998

Keratocyte density and size in conscious humans by digital image analysis of confocal images.

PURPOSE: Confocal microscopy can give images of high magnification and resolution in undisturbed living tissue. It provides new information about the cellular structure of the cornea. Our aim was to measure the density, size and distribution of keratocytes. METHODS: Healthy cornea in four subjects was examined using tandem scanning confocal microscopy. Methods for digital analysis of images were developed. RESULTS: Keratocyte density in confocal cross-sections was greatest immediately under Bowman's membrane (maximum 800 cells/mm2) and decreased sharply towards posterior cornea (minimum 65 cells/mm2). Cross-sectional cell size ranged from 78 to 211 microns2, but did not correlate with depth in the tissue. CONCLUSIONS: Results are consistent with those of earlier work using histological and biochemical techniques in isolated tissue. The methods we have developed enable studies of ongoing processes in conscious humans and can be used to examine diseased tissue as well as the response to injury.

Adult↗
PubMed2004

Objective method of comparing DNA microarray image analysis systems.

Many image analysis systems are available for processing the images produced by laser scanning of DNA microarrays. The image processing system takes pixel-level intensity data and converts it to a set of gene-level expression or copy number summaries that will be used in further analyses. Image analysis systems currently in use differ with regard to the specific algorithms they implement, ease of use, and cost. Thus, it would be desirable to have an objective means of comparing systems. Here we describe a systematic method of comparing image processing results produced by different image analysis systems using a series of replicate microarray experiments. We demonstrate the method with a comparison of cDNA microarray data generated by the UCSF Spot and the GenePix image processing systems.

Algorithms↗
PubMed1988

Modern image analysis methods in hematology.

Image analysis instrumentation presently includes multiparameter microscopy and fast processing. One type of image analysis microscope uses only 1 objective and 2 TV cameras, thereby obtaining 2 magnifications simultaneously: a low magnification for cell selection and a high magnification for further cell analysis. Image analysis is very capable of detecting rare cells among a large population of normal cells. This recognition of very rare events becomes increasingly important in biomedical applications. Investigations such as the detection of mutant cells in the peripheral blood and the detection of rare cancer cells in bone marrow as a sign of early metastases, are described to illustrate the potentialities of image analysis in hematology.

Bone Marrow↗
PubMed1990

Retinal nerve fiber layer analysis by a computerized digital image analysis system.

A computerized digital image analysis system was developed to evaluate the retinal nerve fiber layer (RNFL) by red-free fundus photography. The image intensity of the RNFL was computed for 30 degrees between the disc and macula. Thirty early open-angle glaucoma patients and 30 normal age-matched subjects were examined. The coefficients of variation in the reproducibility of the measurements in normal subjects were 7.3 to 9.9% for the interphotographic studies, and 4.1 to 5.1% for the intraphotographic studies. Using this new image analysis system, the RNFL changes were detected easily and the glaucomatous eyes were distinguished from normal eyes with high sensitivity and specificity, 95% and 80%, respectively. This method will be useful for diagnosis of glaucoma at a very early stage.

Aged↗
PubMed1990

Morphologic studies of sickle erythrocytes by image analysis.

An automated image analysis system was used to characterize the morphology of sickle cells under hypoxic conditions. Images observed by light microscopy were transferred to the image analysis system and were processed to binary (black and white) images, and were then analyzed by area, perimeter, and shape. A circular shape factor (CSF = 4 pi X[area]/[perimeter]2) was found to be useful for elucidating the degree of deformation, but it could not differentiate elongated sickle cells from maple leaf-shaped cells. By combining an elliptical shape factor (ESF = [short axis]/[long axis]) with CSF we could separate deoxygenated homozygous sickle red cells (SS cells) into several morphologically distinct groups, including non-sickled cells, maple leaf-shaped cells, and elongated sickle cells. Using this automated image analysis system, we studied morphologic changes of SS cells exposed to deoxygenation-oxygenation (d-o) cycles between PO2 of 0 and 100 mm Hg (one cycle = 12 minutes) at pHs of 6.9 and 7.4. We found that at both pHs the morphology of sickled cells after the first deoxygenation was predominantly maple leaf-shaped. The number of elongated sickled cells increased as the number of d-o cycles increased, indicating that SS cells changed from maple leaf morphology to classic elongated sickle shape during d-o cycles. Desickling occurred less during the oxygenation phase at pH 6.9 than at pH 7.4 and as the number of d-o cycles increased. These results suggest that during d-o cycles deoxyhemoglobin S fibers may align to form large bundles that do not depolymerize completely even at the arterial oxygen pressure.

Anemia, Sickle Cell↗
PubMed2003

Contaminant source apportionment by PIMMS lead isotope analysis and SEM-image analysis.

By combining scanning electron microscopy (SEM) image analysis and laser ablation plasma ionisation multi-collector mass spectrometry (LA-PIMMS), high precision lead isotope analyses can be obtained from individual metal-rich particles. Soils from Wolverhampton and Nottingham were sampled on the basis of high Pb concentrations or brownfield location. Pressed powder pellets of each were rastered by LA-PIMMS to obtain a bulk Pb-isotope signature. The results plot along an apparent mixing line between the major sources of lead contamination in the UK, that is UK ore deposits and alkyl-lead from petrol additives (Australian ore). Two particularly lead-rich soils were chosen to investigate the lead distribution and isotope variability between size and density fractions. The fine-grained and low-density fractions contained most of the lead and have Pb-isotope ratios comparable with the bulk soils. By contrast, the small, lead-enriched denser fractions contained only a minor proportion of the total lead but Pb-isotope signatures indicating relative enrichment in one or other of the end-members from the mixing line. Further characterisation of individual Pb-rich grains is in progress.

Environmental Monitoring↗
PubMedDate not supplied

Computer image analysis of brain CT images for discriminating hypodense cerebral lesions in children.

A computer software system was designed for the automatic discrimination of focal oedemas from local glioses in brain CT examinations. Image analysis methods were applied to the images of 77 CT examinations of children with focal oedemas (42) or local glioses (35). Textural features derived from the co-occurrence matrix of the lesion's image and a neural network classifier (the multilayer perceptron) were employed for the design of the system. Best classification accuracy (89.6%) was achieved by two textural features (contrast-difference entropy), one hidden layer and three hidden nodes of the classifier. The proposed software system provides new textural information and may be of value to the radiologist in differentiating focal oedemas from local glioses, especially in small lesions, where other radiological criteria are not evident.

Brain↗
PubMed1992

Evaluation of breast carcinoma chemosensitivity by flow cytometric DNA analysis and computer assisted image analysis.

Flow cytometric (FCM) DNA and S-Phase (S%) analyses were compared to computerized image analysis (SAMBA 2005) in 27 breast carcinomas (T3, N0-N1, M0) treated by 3 cycles of preoperative Adriamycin, vincristine, cyclophosphamide, methotrexate, 5-fluorouracil (AVCMF) chemotherapy (CT). Twelve carcinomas had shown objective regression and 15 no regression. Samples studied were obtained by sequential fine-needle cytopunctures. Comparing DNA profiles obtained by both methods before and after the first cycle, it appears that tumors can be divided into 3 groups. In the first group (10 cases), no changes were observed after the first cycle of CT. These tumors before treatment had either single DNA peak without cells in S% and G2M or a major peak with a small S% and G2M peak. The second group (9 cases) showed some changes in DNA profiles with an increased G2M peak but no additional values; these tumors before treatment had a small S% and a G2M peak. In the third group (8 cases), before treatment, all were non-diploid with high S% and high G2M. After the first cycle, all showed obvious changes in DNA profiles with a decrease of the G0/G1 peak and an increased S% and G2M with dispersed additional values along the scale in (G2M) x 2 and (G2M) x 4 regions. When changes were compared to tumor regression in the 1st and 2nd groups, 1/10 and 3/9 cases, respectively, were evaluated as objective regression. In the third group, all had objective regression (p less than 0.001). In most cases, a good correlation was observed with both methods.(ABSTRACT TRUNCATED AT 250 WORDS)

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2002

Protein expression analysis using quantitative fluorescence image analysis on tissue microarray slides.

We have developed a tissue microarray (TMA)-based quantitative fluorescence image analysis (QFIA) method in which protein markers on TMA sections were labeled by immunofluorescence using tyramide signal amplification and a quantitative fluorescence detection system. Using this method, BRCA1 protein expression patterns were studied in the TMA sections of cell lines with known levels of BRCA1 expression and in a small group of human tissue samples obtained from sporadic epithelial ovarian cancers, their corresponding adjacent dysplastic fields, and distant non-tumor areas. We detected distinctive BRCA1 expression patterns in high-grade and low-grade sporadic epithelial ovarian cancers and their associated adjacent dysplastic fields. However, such patterns of expression could not be adequately detected by traditional immunohistochemical staining methods. TMA-QFIA provides a sensitive, automated, and quantitative measurement of protein expression on archived tissue and cell samples and will be a useful tool for protein-level molecular profiling analyses.

BRCA1 Protein↗
PubMed1991

Quantitative DNA analysis in breast carcinomas: a comparison between image analysis and flow cytometry.

We compared the results of quantitative DNA analysis of fresh tumor tissue from 50 invasive breast carcinomas by image analysis and flow cytometry. For image analysis, Feulgen-stained slides of tumor imprints and of disaggregated tumor cytospin preparations were evaluated with the CAS-200 image analyzer. For flow cytometry, propidium iodide-stained disaggregated tumor cells were analyzed with the Coulter EPICS-C flow cytometer. The two methods yielded comparable results. The DNA indices obtained by the two methods showed close correlation by linear regression analysis (r = 0.86, P less than 0.001). There were 26 diploid (52%) and 24 nondiploid (48%) carcinomas. The ploidy pattern between the two methods showed agreement in 41 carcinomas (82%) and discordance in two (4%). Three tumors (6%) were equivocal by flow cytometry and four (8%) by image analysis. The equivocal cases presented potential sources of error in the evaluation of histograms in the near-tetraploid region by flow cytometry and in the near-diploid region by image analysis. Image analysis required smaller tissue samples and permitted direct visualization and selection of tumor cells. It also detected more tetraploid carcinomas. In contrast, flow cytometry analyzed larger cell samples and provided histograms with better resolution. It more readily detected the presence of multiple aneuploid peaks and also the presence of aneuploid peaks in the near-diploid range. The presence of aneuploidy was significantly related to the loss of hormone receptor expression, high mitotic rate, and high histologic and nuclear grades. Our study indicates that image analysis and flow cytometry provide comparable results in a majority of breast carcinomas.(ABSTRACT TRUNCATED AT 250 WORDS)

Breast Neoplasms↗
PubMed1994

Comparative assessment of DNA analysis in effusions by image analysis and flow cytometry.

Cytologic evaluation of body cavity fluids is useful to detect malignancy within the pleural and peritoneal spaces. A definitive diagnosis cannot always be made on cytologic evaluation alone. As malignant processes may show abnormal DNA content, DNA analysis of effusions may be useful. Therefore, we determined the DNA content of 37 effusions by flow cytometry (FC) and image analysis (IA) using the CAS 200. Of the 37 fluids evaluated, 18 were cytologically malignant, 15 benign, and four atypical. Overall, 22 fluids (60%) showed concordance between FC and IA. None of the benign fluids were aneuploid. All showed diploid histograms or diploidy with increased proliferating cells. Three of four atypical fluids had increased proliferating cells by either FC or IA, whereas one was diploid by both methods. Aneuploidy was detected in 13 malignant fluids: five were aneuploid by both methods and eight by only one method. IA identified aneuploidy in five of those eight cases, while three were identified by FC. Three of the cytologically malignant fluids were diploid by both methods, and two showed increased proliferating cells by IA and diploidy by FC. The specificity of both methods was 100%. However, the sensitivity of identifying a malignant fluid by aneuploidy is low, 44% for FC and 55% for IA. IA appears to identify small aneuploid populations more frequently than FC. The detection of aneuploidy in effusions is highly suggestive of malignancy, and the combination of both techniques gives the highest detection rate (72%). However, neither are as sensitive as traditional cytologic evaluation with the occasional use of additional histochemical stains.

DNA, Neoplasm↗