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PubMed2002

An automatic block and spot indexing with k-nearest neighbors graph for microarray image analysis.

MOTIVATION: In this paper, we propose a fully automatic block and spot indexing algorithm for microarray image analysis. A microarray is a device which enables a parallel experiment of ten to hundreds of thousands of test genes in order to measure gene expression. Due to this huge size of experimental data, automated image analysis is gaining importance in microarray image processing systems. Currently, most of the automated microarray image processing systems require manual block indexing and, in some cases, spot indexing. If the microarray image is large and contains a lot of noise, it is very troublesome work. In this paper, we show it is possible to locate the addresses of blocks and spots by applying the Nearest Neighbors Graph Model. Also, we propose an analytic model for the feasibility of block addressing. Our analytic model is validated by a large body of experimental results. RESULTS: We demonstrate the features of automatic block detection, automatic spot addressing, and correction of the distortion and skewedness of each microarray image.

Algorithms↗
PubMed2003

Cytometrical image analysis for immunohistochemical hormone receptor status in breast carcinomas.

A cytometrical image analyzing method for nuclear protein was established using WinROOF, a commercially available, inexpensive software, to determine the status of both estrogen and progesterone receptors. Immunohistochemical evaluation of estrogen receptors (ER) and progesterone receptors (PR) was performed with the anti-ER (clone 1D5) and the anti-PR (clone PgR636), respectively, combined with dextran polymer reagent EnVision+, all of which are approved in vitro diagnostics in Japan. The immunostained results were captured as digital images in Windows, and then analyzed in WinROOF with macroinstructions for analyzing each captured area either immunolabeled with chromogen or counterstained with hematoxylin. This image analysis method graded the immunostained nuclei of carcinoma cells based on staining intensities, and calculated the labeling index (LI) for both ER and PR. Furthermore, the LI correlated highly with the results from a histology score (HSCORE) when 20 breast carcinomas were quantified. Regarding ER, when 20% in the LI was considered as the cut-off point for positive, the positivity of ER in computer-assisted analysis was 75% (15 of 20 cases), and was completely concordant with that of HSCORE-based analysis. These results indicate that the cytometrical image analysis-based quantification could be appropriately applied to the objective determination of the immunohistochemical status of both ER and PR.

Biomarkers, Tumor↗
PubMed1996

Interlaboratory comparison of DNA image analysis.

Interlaboratory quality assurance studies have been conducted for DNA flow cytometry, but not for DNA image analysis systems. The purpose of this study was to investigate if concordance of DNA image analysis systems existed with respect to classification and staining of standardized material. In three separate rounds, human liver cells were measured randomly by means of the image cytometry system present in each participating laboratory. The features integrated optical density (IOD) and AREA were reported. The relationship between the coefficient of variation (CV) of the 2c and 4c peak were compared with three models. In the three rounds the number of participating laboratories was 11, 14 and 11, respectively. Sequential plotting of normalized IOD values yielded useful information about intra-measurement variation. Comparison of measurements in specimens stained in the participating and central laboratory revealed similar CV values. In general, the precision of the instruments, expressed as the 4c/2c and 8c/2c ratios was good. The accuracy of the different laboratories expressed as the CV of IOD for the three rounds varied from 2-17%. The relation of the CVs of the 2c and the 4c peaks was best fit with the model of the addition of two normal distributions. We conclude that interlaboratory comparison of DNA measurements performed on different instruments is certainly feasible and could facilitate improvement in quality standards.

DNA↗
PubMed1990

Quantitative evaluation of melanoma cell invasion in three-dimensional confrontation cultures in vitro using automated image analysis.

Tumor invasion is a crucial feature of tumor growth in vivo. Confrontation cultures of multicellular melanoma spheroids and embryonic chick heart fragments provide a model for invasive growth in vitro. We have developed an image analysis method, which facilitates the objective measurement of tumor cell invasion in this model. Cryostat sections of confrontation cultures were immunohistochemically stained with an antiserum directed against the stromal component for automated recognition of the stroma tissue. The slides were automatically processed by a grey level based computerized image analysis system. On Spearman's rank correlation test, 25 out of 39 parameters correlated with the reference value of invasion, which was derived from the subjective evaluation of five independent observers. Two parameters combining the stroma margin and the total amount of stroma tissue completely reproduced the judgement of the morphologists in our test set. The quantitative evaluation of tumor invasion in vitro by automated image analysis may be helpful in pharmacologic and pathogenetic studies of tumor growth.

Animals↗
PubMed1994

Computer-aided image analysis applied to immunogold-silver staining: evaluation of proliferating cell nuclear antigen (PCNA)-reactive sites in paraffin sections.

Feasibility of the combination of the immunogold-silver staining method (IGSS) and computer-aided image analysis was assessed for the detection of antigen in an immunostained, paraffin-embedded section. Using low-temperature IGSS, we stained a specimen of human oral squamous cell carcinoma with a monoclonal antibody, PC 10, against a proliferating cell nuclear antigen (PCNA/cyclin), and the section was analyzed by ACAS 570 interactive laser cytometry. The PCNA-positive cells, exhibiting a heteromorphic texture, were contrasted by the dark staining of their nuclei, but showed heterogeneity in staining intensity from cell to cell. Using a conventional microscope light source rather than a laser, and by employing the COMPLEMENT DATA program (which permits inversion of the data values) installed in the ACAS 570 software system, we were able to obtain a 'complemental image' which replicated the real immunohisto-morphology. Approximately 30-35 cells from three different areas in the same section were selected by DEFINE CELL and MARK AREA programs, and quantitative image analysis was performed in terms of cell integrated value, area, perimeter, and shape factor indicated in histogram form. The combined utilization of IGSS with computer-aided image analysis was demonstrated to offer a crucial advantage for the quantitative assessment of immunostained sections.

Humans↗
PubMed2000

Objective quantification of plaque using digital image analysis.

Dental plaque is the precursor to many oral diseases (e.g. gingivitis, periodontitis, caries) and thus its removal and control are an important aspect of oral hygiene. Many of the oral care products available today remove or inhibit the growth of dental plaque. Historically, the antiplaque efficacy of products was measured in blinded clinical trials where the amount of plaque on teeth is assessed via subjective visual grading with predefined scales such as the Turesky index. The ability of the examiner to consistently apply the index over time and the sensitivity of the scales often leads to large, expensive clinical trials. The present invention is an automatic measurement of plaque coverage on the facial surfaces of teeth using a digital image analysis technique. Dental plaque disclosed with fluorescein is digitally imaged under long-wave ultraviolet light. Ultraviolet illumination of fluorescein-disclosed plaque produces an image where the pixels of the image can be categorically classified based on color into one of five classes: teeth; plaque; gingiva; plaque on gingiva, or lip retractors. The amount of plaque on teeth can be determined by summation of the number of plaque pixels. The percent coverage is calculated from the number of plaque pixels and teeth pixels in the image. The digital image analysis of plaque allows facial plaque levels to be precisely measured (RSD = 3.77%). In application, the digital image analysis of plaque is capable of measuring highly significant plaque growth inhibition of a stannous fluoride dentifrice with as few as 10 subjects in a cross-over design.

Analysis of Variance↗
PubMed1992

Quantitative evaluation of nuclear cataract using image analysis.

To quantitatively evaluate nuclear lens opacification, we applied image analysis techniques. Utilizing a newly developed anterior eye segment analysis system, Scheimpflug slit images were taken in 65 eyes with transparent lenses and 31 eyes with nuclear cataract. In transparent lenses, scattering light intensity of the anterior fetal nucleus (AFN) was equal to or less than that of the posterior fetal nucleus (PFN). In eyes with nuclear cataract, scattering light intensity of the AFN was higher than that of the PFN. Utilizing this phenomenon, nuclear opacification was evaluated by the difference of scattering light intensity between the AFN and PFN (delta i value). The methodology applied in this investigation provides precise grading of nuclear cataract and progression thereof.

Adult↗
PubMed1990

Image analysis of the chromatin organization in the nuclear domains of freeze fractured hepatocytes and lymphocytes.

The complex organization of the interphase nucleus can be analyzed, by way of thin sectioning and also freeze-fracture. This approach has previously been utilized in association with image analysis to quantitatively describe the organization of isolated rat liver nuclei and nuclear matrices. The main nuclear domains which, in section, present marked differences due to their electron-density, can be identified in replicas with more complex procedures, based on the quantitative evaluation of the number of particles per unit area and mainly by using image analysis. A quantitative analysis of the nuclear substructures has been performed by way of image analysis on in situ nuclei of freeze-fractured cells presenting marked differences in the heterochromatin quantity, such as hepatocytes and lymphocytes. The replicated nuclear particles have been classified according to their diameter and the obtained histograms have been quantitatively evaluated. The nuclear domains, heterochromatin, interchromatin, nucleolus, present characteristic ratios among the three main classes of particles; that is, ribonucleoproteins, solenoid filaments and solenoid fibre aggregates. The typical patterns of the nuclear domains can be further stressed by selecting a single class of particles and by examining its topographic localization. While interchromatin and nucleolar domains present a similar quantitative pattern in hepatocytes and lymphocytes, the heterochromatin of lymphocytes contains a significative higher percentage of solenoid aggregates than that of hepatocytes.

Animals↗
PubMed1992

[Progress on chromosome image analysis systems].

This is a review of the development of automated systems of chromosome analysis. The history of its development reflects the progress of image analysis systems and that of method of chromosome analysis in man. It can be chronologically summarized as follows. Period I: 1960s-early 1970s when chromosome banding methods were discovered. Period II: early 1970s-early 1980s when the first image analysis system of chromosome was commercialized. Period III: early 1980s-present. Research and development of the automated analysis system of radiation induced chromosome aberrations, conducted under the national project of cross-over research on underlying technology of nuclear energy, Science and Technology Agency, Japan, is briefly reported.

Chromosomes↗
PubMed1994

Detection of discrete androgen receptor epitopes in prostate cancer by immunostaining: measurement by color video image analysis.

To determine whether multiple features of immunohistochemical staining of the androgen receptor (AR) in prostate cancer could reliably predict androgen dependence, tumor biopsy specimens from 30 patients (stages A-D2) were stained using anti-peptide antibodies to the amino- and carboxyl-terminal of the AR. Measurements were made of the mean area and total amount (i.e., integrated optical density) of AR staining in at least 20 fields per section using a color video image analysis system, and the mean intensity of AR staining per cell and the percentage of AR positive tumor cells were derived. Video image analysis measurement identified quantitative differences in AR staining between the two antibodies, suggesting that this approach may provide a means of identifying receptor variants in prostate tumors. The AR staining measurements were analyzed by discriminant function analysis to assign individual cases to good and poor clinical outcome groups. AR staining features measured with a single antibody (e.g., amino-terminal) were sufficient to predict outcome following hormonal therapy in stage D2 patients (predictive value, 1.0), whereas all features of AR staining measured with both antibodies were required for the entire patient group (predictive value, 0.97). The principal discriminant in both patient groups contributing to the correct assignment of outcome was the mean intensity of AR staining per cell. These findings suggest that AR staining features measured by video image analysis have the potential to predict outcome in prostate cancer.

Aged↗
PubMed1993

Quantitative morphology and shape classification of neurons by computerized image analysis.

We describe a new image processing method for semiautomatic quantitative analysis of neuronal morphology. It has been developed in a specific image analysis environment (IBAS 2.0), but the algorithms and the methods can be employed elsewhere. The program is versatile and allows the analysis of histological preparations of different quality on the basis of different levels of evaluation and image extraction. Some significant algorithms have been implemented (i.e. one for multiple focus image acquisition and one for automatic cell body shape recognition and classification). A wide set of specific morphological parameters has been defined to allow a better mathematical characterization of neuronal morphology as regards both dendrite trees and cell bodies. Cell bodies' shapes can be classified automatically, defining different neuronal populations. This is done by evaluating the number of main dendrites and perikarya shapes through a multi-valued-decision-tree based method, tested on somatostatin-positive cells in mouse brain. The methods presented have been applied to analysis of neurons, but they can well be used for any quantitative morphological study of other cell populations.

Algorithms↗
PubMed1999

Cytologic score and DNA-image analysis in the classification of borderline breast lesions: a prospective study on 47 fine-needle aspirates.

We prospectively evaluated the accuracy of the cytologic score system developed by Masood et al. combined with DNA-image analysis in the subclassification of 47 fine-needle aspiration samples with cytologic features of borderline breast lesions. Cytologic scores ranged between 12-18. All cases underwent surgical excision of the lesion, and histology revealed 24 cases of florid hyperplasia, 8 of atypical hyperplasia, and 11 noninvasive and 4 invasive ductal carcinomas. DNA-image analysis demonstrated 33 diploid and 14 aneuploid cases. Diploid samples were divided into slowly proliferating (S + G2/M < or = 13%) and rapidly proliferating (S + G2/M > 13%) cases. By considering florid hyperplasia a "low-risk" lesion and by amalgamating atypical hyperplasia, and in situ and invasive ductal carcinoma in the category of "high-risk" lesions, the positive predictive value of a score value > 16 was 100%. In cases scoring < or = 16, the slowly proliferating pattern had a negative predictive value of 95%, while the aneuploid and rapidly proliferating patterns had a positive predictive value of 100% and 63%, respectively. We conclude that a combination of cytologic score evaluation and DNA-image analysis is very useful in differentiating "low-risk" from "high-risk" cases in the field of breast borderline lesions, thus improving the impact of fine-needle aspiration diagnosis on patient management.

Biopsy, Needle↗
PubMed1991

Computer-assisted image analysis of nucleolar organizer regions (NORs): a pilot study of astrocytomas and glioblastomas.

Computer-assisted image analysis was used to measure nucleolar organizer regions (NORs) of 22 astrocytomas and glioblastomas. Image analysis provides reproducible information about number and size of NORs together with further karyometric data, which can be compared and processed with other patient-related data. Our study exhibits a statistical relationship between number and size of NORs and malignancy of the measured gliomas.

Astrocytoma↗
PubMed1994

In vitro quantification by image analysis of inotropic and chronotropic effects of drugs on cultures of cardiac myocytes.

Quantitative image analysis is used to measure the inotropic and chronotropic effects of drugs on cultured heart cells maintained at 37 degrees C on the stage of an inverted light microscope, and sequentially superfused with control and treatment media. The beating of the cardiac myocytes is evaluated by simultaneously selecting up to eight areas, including cell edges, from digitized video image. The sizes and positions of these areas are controlled by the operator. To analyze the motion of cell edges in each area, the computer measures the shift of the mass center of pixels' grey levels. Finally, a few parameters are calculated for the eight areas and displayed graphically. In order to assess treatment effects, appropriate statistical tests are performed on the data. Image analysis is an efficient screening test for evaluating the pharmacologic or toxic effects of a substance on isolated or cultured cardiac myocytes from various species.

Animals↗
PubMed1992

Mast cell quantitation by image analysis in adult mastocytosis and inflammatory skin disorders.

Mast cell numbers were quantitated in adult cases of mastocytosis demonstrating non-diffuse perivascular and upper dermal concentrations of mast cells. Using the Leder stain and computerised video image analysis, a mean of 382 (+/- 28 SE) mast cell per mm2 were counted in the superficial dermis in skin biopsies from 30 adult cases of mastocytosis, in contrast to a mean of 43 (+/- 5 SE) mast cells per mm2 in skin biopsies from 50 inflammatory dermatoses represented by subacute dermatitis, pigmented purpuric dermatosis, erythema multiforme, lichen planus and granuloma annulare. Ten skin biopsies showing no significant inflammation had a mean of 54 (+/- 7 SE) mast cells per mm2 in the upper dermis. The mean area of individual mast cells as assessed by image analysis in the mastocytosis group was 47.40 microns 2 (+/- 2.26 microns 2, SE) which was significantly different (P < 0.01) than the mast cell area (32.34 microns 2 +/- 2.22 microns 2, SE) in all other groups combined. Computerised video image analysis represents an alternative technique which is useful in assessing mast cell numbers and particularly mast cell size in adult cases of macular mastocytosis and in other dermatoses.

Adult↗
PubMed1993

Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ.

We extend use of the lacZ reporter gene for tumor biology. Intracerebral growth of 9L/lacZ, a gliosarcoma cell line that stably expresses lacZ, was evaluated in syngeneic rats. The reporter gene product, Escherichia coli-derived beta-galactosidase (beta-gal), was detected histochemically on tissue sections. This permits visualization of disseminated tumor and, as shown here, facilitates image analysis. We show that the beta-gal marker protein itself can serve as a tumor antigen in appropriate contexts. Quantitative image analysis of tumor areas is used to show that immunization with beta-gal protects against tumor growth. Abnormal beta-gal- areas are easily detected, facilitating study of antigenic modulation. The tumor studied did not escape through this mechanism. All abnormal beta-gal- areas examined were shown to reflect accumulation of inflammatory or reactive cells, not tumor. Taken together, these findings show several ways in which the lacZ reporter gene can be exploited to facilitate quantitative analysis of disseminated tumor growth within the brain. They draw attention to the growing appreciation that tumor antigens need not be cell surface molecules.

Animals↗
PubMed1992

[An experimental study on the wear of HDP socket in total hip arthroplasty--quantification of worn surface by SEM 3-D image analysis and effect of gamma-ray-irradiation of HDP on improving tolerance to wear].

In order to evaluate the usefulness of gamma-ray-irradiation to improve the tolerance to wear of the sockets, the worn surface of the 2.5 M rad gamma-ray-irradiated HDP sockets after total hip arthroplasty has been quantified by a newly-developed 3 dimensional (3-D) image analysis method in combination with scanning electron microscopy (SEM). The SEM 3-D image analysis revealed marked deformation, consisting of unevenness with about a 20-micron difference in height, of the worn surface of the ordinary HDP socket, while the same analysis revealed a smooth surface in the gamma-ray-irradiated HDP socket. The decrease in thickness of the gamma-ray-irradiated HDP socket was about 1/4 that of the ordinary HDP socket. These results indicate that SEM 3-D image analysis is capable of quantifying the morphology of the worn surface and that gamma-ray-irradiation improves the tolerance to wear of the socket, thus making its surface resistant to deformation.

Animals↗
PubMed2002

Color image analysis of cervical neoplasia using RGB computer color specification.

OBJECTIVE: To establish quantitative color image analysis for cytology, red, green and blue (RGB) color specification was applied to Papanicolaou-stained cervical smears. STUDY DESIGN: Cell samples used in this study was those from 300 cervical specimens. We analyzed the color tone of nuclei and cytoplasm of the squamous cells in the cervical smear by means of computer image analysis. RESULTS: Papanicolaou stained nuclei displayed basophilic blue to purple. When they were hyperchromatic and deeply stained, B and G values decreased in value. The RGB values of cytoplasm and nuclei decreased significantly (P < .01) as their degree of cellular atypia increased. CONCLUSION: Using RGB color specification to analyze Papanicolaou-stained cervical smears, a significant difference was perceived in the nucleus and cytoplasm between different groups of squamous cells, from normal, dysplastic and squamous cell carcinoma. These findings may help to establish automated cytology.

Carcinoma in Situ↗