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PubMed1988

The evaluation of estrogen receptor in primary breast carcinoma by computer-assisted image analysis.

A monoclonal antibody prepared against estrogen receptor has been shown to be specific and sensitive for the detection of estrogen receptor in human breast lesions by use of immunohistochemical methods. Two hundred selected cases of primary breast carcinoma were assayed for estrogen receptor content by biochemical and immunohistochemical procedures. Quantitative evaluation was by biochemical, immunohistochemical, and automated computer-assisted image analysis using the Cell Analysis System's CAS/100 machine (Lombard, IL). Quantitative estrogen receptor content was determined by dextran-coated charcoal analysis and sucrose density gradient analysis. Immunohistochemical evaluation incorporated both intensity and distribution of staining, yielding a subjective score, histologic score (HSCORE). An objective quantitation, also incorporating intensity and distribution of staining, was done by computer-assisted image analysis, quantitative immunocytochemical score (QIC SCORE). HSCORE analysis was done with and without methyl green counterstain with no loss of sensitivity. Comparison of QIC SCORE with the biochemical and immunohistochemical analysis of the tissues examined revealed excellent sensitivities and specificities. These data suggest that automated image analysis provides an effective qualitative and quantitative means of evaluating estrogen receptor content in human breast cancers.

Antibodies, Monoclonal↗
PubMed1994

Computerized image analysis and flow cytometric evaluation of ovarian borderline tumors: a study of 24 cases.

DNA content evaluation in the study of ovarian borderline tumors has been shown to be a useful adjunct to histopathologic diagnosis. This study compares flow cytometry and computerized image analysis (CIA) in evaluating the DNA content of these lesions. Twenty-four cases of ovarian borderline tumors (15 serous, 9 mucinous) were studied utilizing formalin-fixed paraffin-embedded tissue. Flow cytometry of cell suspensions and CIA of cell suspensions and paraffin sections were compared in the evaluation of DNA content. Twenty-three tumors (96%) were diploid and only 1 (4%) was aneuploid. There was 100% correlation between flow cytometry and CIA of cell suspensions. Image analysis of Feulgen-stained paraffin sections was found to be unreliable when compared with the use of cell suspensions in the evaluation of DNA content. Clinical follow-up in 17 patients showed recurrent disease in two patients with diploid tumors and no evidence of disease in the patient with an aneuploid tumor. The reported rate of aneuploidy in borderline tumors varies depending on the diagnostic method employed in evaluating DNA content. This study shows good correlation between flow cytometry and image analysis of cell suspensions and supports the low rate of aneuploidy found in these lesions by other authors utilizing these methods. The majority of borderline ovarian tumors have been shown to have a diploid DNA content; however, the finding of aneuploidy in histologically classified borderline tumors warrants close clinical follow-up since aneuploidy in these tumors has been associated with an adverse prognosis.

Aneuploidy↗
PubMed1996

Detection of structural damage from glaucoma with confocal laser image analysis.

PURPOSE: To determine which structural optic nerve head parameters measured with confocal scanning laser image analysis that best discriminate between normal persons and those with glaucoma. METHODS: One randomly selected eye of 53 patients with early open-angle glaucoma (average visual field mean deviation = -4.8 dB) and of 43 age-, race-, and refractive error-matched normal subjects were studied. The performance of nine structural measures was evaluated with linear multivariate analysis and a neural network: cup area, cup to disc area ratio, rim area, height variation contour, cup volume, rim volume, cup shape measure, mean retinal nerve fiber layer thickness, and retinal nerve fiber layer cross-section area. A discriminant function was derived with two thirds of the sample and its discriminant power tested on the remaining one third. This was repeated twice so that the entire sample was used for training and testing. A neural network was trained and tested in the same way. Stereoscopic color optic nerve photographs of the same eyes were evaluated qualitatively by three experienced, masked observers. Receiver operating characteristic (ROC) curves of discriminant function, neural network results, and qualitative evaluation were plotted. Comparisons of the areas under the ROC curves were performed with nonparametric statistics. RESULTS: There were statistically significant differences between the normal and glaucoma groups for all measures (P < or = 0.007) except for height variation contour, mean retinal nerve fiber layer thickness, and retinal nerve fiber layer cross-section area. Cup shape measure provided the single best measure to distinguish between normal subjects and those with early glaucoma and had a diagnostic precision of 84%. Neural network diagnostic precision, when all measures were used, was 92% and decreased to 82% when cup shape measure was omitted. The area under the ROC curve when all measures were combined was 0.94; it was significantly lower (P = 0.04) when cup shape measure was omitted (area = 0.84). The area under the ROC curve for qualitative optic disc evaluation by experienced observers was 0.93. There was no statistically significant difference between qualitative evaluation and neural network performance (P = 0.80). CONCLUSIONS: Cup shape measure, the statistical third moment of the distribution of depth values of the optic nerve head obtained with confocal laser image analysis, can be used to discriminate between normal persons and those with early glaucomatous damage with high diagnostic precision.

Female↗
PubMed1993

Image analysis in the evaluation of biomaterials.

An examination regime, based on a computer-aided image analysis system, has been developed for the quantitative evaluation of the local tissue response to biomaterials. This procedure involves the immunoenzymic staining of tissue sections using monoclonal antibodies specific for certain inflammatory cell types. An avidin-biotin-horseradish peroxidase staining method is used to identify antibody binding sites and the sections are assayed using a computer-aided image analysis system. This regime facilitates the rapid and accurate measurement of 30 cell related parameters in sections stained for macrophages, polymorphonuclear leucocytes, and other cells.

Animals↗
PubMedDate not supplied

Quantitative evaluation by image analysis of platelet morphological modifications after contact with polyvinylacetate.

BACKGROUND: In order to investigate platelet activation after contact with artificial materials, which is an important aspect of biocompatibility especially for the blood-contacting devices, platelet morphological modifications and spread area were evaluated by light microscopy and image analysis after contact with glow discharge-treated polybutylene terephthalate coated with a polymer for platelet concentrate filtration. METHODS: A hydrophilic polymer made of partially hydrolyzed polyvinyl-acetate containing polyethylene oxide/poly-propylene oxide copolymer block as lateral chains (PVA) (Biofil S.r.l., Cavezzo, Modena, Italy) was evaluated. After contact with PVA, platelets were allowed to settle on a siliconized slide and then fixed and stained. The specimens were analyzed by image analysis. The percentages of spreading, round and dendritic shapes, as well as the presence of aggregates, were evaluated, and the mean area of the spread platelets was measured. RESULTS: PVA induced significant variations neither in the percentages of shape change distribution, nor of the mean spread area. However it determined a statistically significant reduction in platelets with the area from 60 to 70 mu 2. Such minimal variations agree with the results we obtained in the past, namely a non significant platelet adhesion induced by the same material. CONCLUSIONS: The method confirms the results of platelet adhesion and release reaction (the study of release reaction needs more refined but more expensive methods). However, the study of morphological modifications by image analysis is not suitable for testing materials that induce massive platelet adhesion, because the number of the residual platelets could be too low for the microscopic evaluation.

Analysis of Variance↗
PubMed2004

Imaging analysis of STAT1 and NF-kappaB translocation in dendritic cells at the single cell level.

Rapid assessment of immune or stem cells, which are now widely applied in the clinical setting of cancer treatment, is necessary to speed their development and to determine their quality. We have evaluated immature dendritic cells (iDC) by semiautomated imaging cytometry which provides detailed assessment at a single cell level. Nuclear translocation of NF-kappaB was studied by imaging analysis as well as electrophoretic mobility shift assay with an excellent correlation (r=0.981) over a broad range of lipopolysaccharide (LPS) concentrations. Imaging analysis was time saving (5 h vs. 3 days), and required 30- to 100-fold less cells per analysis. Single cell information revealed remarkable heterogeneity between individual iDC and permitted detection of responses to 40 pg/ml of LPS. In IL-1beta/IFNgamma activated iDC, STAT1 responses preceded NF-kappaB responses, and the expression of both was strongly correlated in individual cells (p<0.001). IFNgamma amplified IL-1-induced NF-kappaB responses. NF-kappaB responses to IL-1beta, CD40L, and LPS were donor-dependent (n=7), correlated with the quality of iDC preparations (p=0.002), and IL-12 p70 production (p=0.010). NF-kappaB measurements in iDC within mixed cell cultures (iDC, NK, K562) demonstrated that these strategies are applicable for analyses of complex cell-cell interactions. Imaging analysis is a method that could be valuable for quality control of cell therapy preparations.

Antineoplastic Agents↗
PubMed2005

Determination of gunshot residues with image analysis: an experimental study.

In firearm injuries, assessment of the firing range and determination of entrance and exit wounds are important. For this reason, evaluation of the amount and distribution of gunshot residues (GSRs) is necessary. Several methods and techniques for GSR analysis have been developed. Although these methods are relatively sensitive and specific, they may require expensive dedicated equipment. Therefore, a simple, easily applicable, more convenient method is needed. A total of 40 experimental shots were made to calf skin from distances of 0, 2.5, 5, 10, 20, 30, 45, and 60 cm. Eighty samples were taken from the right and left sides of the wounds, and Alizarin Red S dye staining was performed. The amounts of GSR particles were measured with image analysis. GSRs were detected in all shots. The mean size of the distribution area of barium and lead elements around the wound had a significant negative correlation with increasing shooting distance (r = -0.97, p < 0.001). As the distance increased, the amount of GSR decreased, and this decrease rate was nonlinear. Variance analysis suggested significant differences between data groups depending on range (p < 0.001). The image analysis method may solve some of the standardization problems for evaluation of GSRs. GSR detection with the image analysis method does not require experienced personnel and may be a suitable method for scientific studies and for routine purposes.

Animals↗
PubMed1984

[Results of microscopic image analysis of the thyroid gland fine needle biopsies].

In previous cytomorphometrical studies applying a microscopic image analysis system it was shown that follicular adenomas and highly differentiated follicular carcinomas of the thyroid gland can be distinguished. Basing on these results, the usefulness of the method was examined in selected cases of routine materials. 143 fine needle biopsies were analysed by means of the image analysis system. Moreover, in 52 cases the results could be checked by histological investigations of surgical samples. In comparison with the cytomorphological findings, the results of the cytomorphometrical analysis demonstrate a higher grade of assurance of the diagnostic statements. It was confirmed, that it is possible to diagnose highly differentiated follicular carcinomas of the thyroid gland with the method demonstrated above.

Adenocarcinoma↗
PubMed2005

Semi-automated digital image analysis of patellofemoral joint space width from lateral knee radiographs.

OBJECTIVE: To design a semi-automated program to measure minimum patellofemoral joint space width (JSW) using standing lateral view radiographs. DESIGN: Lateral patellofemoral knee radiographs were obtained from 35 asymptomatic subjects. The radiographs were analyzed to report both the repeatability of the image analysis program and the reproducibility of JSW measurements within a 2 week period. The results were also compared with manual measurements done by an experienced musculoskeletal radiologist. RESULTS: The image analysis program was shown to have an excellent coefficient of repeatability of 0.18 and 0.23 mm for intra- and inter-observer measurements respectively. The manual method measured a greater minimum JSW than the automated method. Reproducibility between days was comparable to other published results, but was less satisfactory for both manual and semi-automated measurements. The image analysis program had an inter-day coefficient of repeatability of 1.24 mm, which was lower than 1.66 mm for the manual method. CONCLUSIONS: A repeatable semi-automated method for measurement of the patellofemoral JSW from radiographs has been developed. The method is more accurate than manual measurements. However, the between-day reproducibility is higher than the intra-day reproducibility. Further investigation of the protocol for obtaining sequential lateral knee radiographs is needed in order to reduce the between-day variability.

Adult↗
PubMed1988

A versatile video image analysis system for microcirculatory research.

A modular image analysis system is presented consisting of a personal computer equipped with a real time video digitizer, an interactive control unit and a graphic tablet. Together with the corresponding software modules this system can be used for a number of image processing procedures in microcirculatory research including image enhancement, measurement of morphological parameters and image brightness as well as determination of vessel diameter and blood flow velocity.

Animals↗
PubMed1996

The adherence of oral isolates of Enterobacteriaceae to HeLa cells. An in vitro method using image analysis.

An in vitro model and an image analysis were designed to improve on existing quantification methods in the assessment of the adherence of Enterobacteriaceae to human epithelial cell monolayers. Adherence to HeLa cell monolayers of three oral isolates and one type strain from each of four species of Enterobacteriaceae over two incubation time periods was examined. Correction for actual cell area and a cube root transformation of the data to stabilize variance were applied. While behaviour varied between strains within species, E. cloacae was the most, and K. pneumoniae the least, adherent species irrespective of the incubation period. Increasing the incubation period from 30 min to 60 min resulted in greater adherence for E. cloacae, E. coli, and C. freundii, but not K. pneumoniae strains. The method permits the reliable measurement and valid analysis of the adherence of Enterobacteriaceae to cultured epithelial cell monolayers.

Cell Adhesion↗
PubMed1999

Quantitative image analysis of F-actin in endothelial cells.

OBJECTIVE: Filamentous actin (F-actin) plays a central role in maintaining endothelial barrier function. Thrombin and histamine, two inflammatory mediators that increase endothelial permeability, can alter F-actin production and distribution. In this study, we use a newly developed image analysis technique to show that these two inflammatory mediators differentially alter F-actin structure. METHODS: Human umbilical vein endothelial cells were grown to confluence and treated with either histamine (1 microM), thrombin (1 microM) or the agonist's vehicle. The endothelium was stained with BODIPY-phallodin, and digitized images were taken of the treated cells. The digitized images of individual human umbilical vein endothelial cells (HUVEC) were imported into a F-actin image analysis program (FAAP) and converted to layers, each one pixel thick. The program then determined the mean gray level (which corresponded to the amount of F-actin) in each layer starting from the outside of the cell (layer 1) and progressing in one pixel layer increments towards the center of the cell (layer 32). RESULTS: Both inflammatory mediators increased endothelial F-actin production, however, the distribution of the actin was different. Thrombin increased the presence of stress fibers, while also decreasing peripheral banding actin. In contrast, histamine had no effect on peripheral actin compared to control, but did increase the presence of F-actin stress fibers. CONCLUSIONS: These results establish that thrombin and histamine alter endothelial F-actin production in different locations within the cell, which can be quantified using an image analysis program.

Actins↗
PubMed1997

Testis biopsy image analysis effectively quantifies spermatogenic cell types.

PURPOSE: Evaluation of the testis biopsy specimen in male infertility has been hampered by the qualitative rather than quantitative nature of routine histological techniques and interpretation. Although useful, flow cytometry has demonstrated significant limitations due to its inability to differentiate among haploid cell types, spermatozoa and spermatids. Thus, deoxyribonucleic acid image analysis was performed on testis biopsy specimens to combine the advantages of quantitative ploidy and morphological characteristics to develop a more effective quantitative system. MATERIALS AND METHODS: Image analysis using a specifically designed filter was performed on Feulgen stained 5 microns. sections of paraffin embedded testicular tissue. The archival testicular tissue had been obtained using standard biopsy techniques from patients with azoospermia and normal spermatogenesis on routine hematoxylin and eosin processed tissue. RESULTS: The overall findings (mean plus or minus standard deviation) in 20 biopsies from 18 men are haploid 54.2 +/- 5.6, diploid 31.2 +/- 5.2 and tetraploid 15.0 +/- 4.6%. Spermatozoa and spermatids contributed 23.2 +/- 5.7 and 30.9 +/- 7.6% to the total cell content, respectively. A mean of 2,462 +/- 254 cells were counted per case. CONCLUSIONS: We used image analysis to evaluate testicular biopsies. This technique can quantitatively determine the percentages of various cell types within the seminiferous tubules. Furthermore, it can differentiate between spermatozoa and spermatids based on morphological characteristics. This technique, which combines ploidy and cell morphology characteristics, represents a significant advancement in the interpretation of the testicular biopsy, and has implications for redefining the presently used system of qualitative interpretative assessment.

Adult↗
PubMed1999

Automated identification of stained cells in tissue sections using digital image analysis.

OBJECTIVE: To develop a novel automated image analysis system to differentiate immunohistochemically stained cells from background. STUDY DESIGN: Cell segmentation was performed by applying global thresholding algorithms to find an approximate threshold at which cells could be separated from background followed by a novel refinement algorithm to erode edge pixels of the region. To separate overlapping cells, a new decomposition method was developed that uses both semantic knowledge and high-level relational information. Both the cell segmentation and separation methods were evaluated on images of stained tissue sections and the manually outlined cell areas and numbers compared to the computed. RESULTS: Macrophage areas computed at the first stage by Otsu's algorithm did not differ significantly (P = .07) from those traced manually, while the areas computed by Kittler's and Kurita's algorithms did not agree (P < .01). Both Otsu's and Kurita's algorithms performed well when combined with edge pixel erosion. Kittler's algorithm proved unsuccessful even with edge erosion. Comparison of the computed and manually determined cell numbers showed a significant correlation, and regression analysis resulted in the unity curve. CONCLUSION: A combination of global thresholding and a novel edge erosion technique allowed identification of immunohistochemically stained macrophages; the computed cell areas agreed with the manual results.

Aorta↗
PubMed1992

[Image analysis of precancerous and carcinomatous changes of large intestine in histopathologic diagnosis].

This article reports the image analysis in the study of normal mucosa, polypoid adenoma, villus adenoma, atypical hyperplasia and tubular adenocarcinoma of the large intestine (15 cases each). The following 8 parameters: nuclear area (NA), plasma area (PA), ratio of nuclear to plasma area (RNPA), mean nuclear diameter, nuclear length, nuclear width, cell volume density and cell surface area density (SV) were observed and compared. Each value was tested by stepwise discriminational classification, giving an accuracy rate of 98%. These results suggested that analysis of variance in these groups was very significant in statistics (P less than 0.01). The mean values of RNPA and SV were especially significantly different between the adenocarcinoma group and the other groups (P less than 0.01). These results show that image analysis may be of importance in the histopathologic differentiation of precancerous from carcinomatous lesions.

Adenocarcinoma↗
PubMed2002

[Research on slit lamp bio-microscope image analysis system].

Slit lamp bio-microscope image analysis system can provide objective and quantitative diagnosis evidences for ophthalmologists and make available efficient storing, management and intercourse of sufferer information and diagnosis results. In this paper, the system is discussed on its devices, software structure, analysis process and algorithm.

Algorithms↗
PubMed2000

Comparative image analysis of EGF immunoreaction in rat submandibular gland using 3,3'-diaminobenzidine with metal enhancer substrate.

We have shown the efficacy of image analysis using 3,3'-diaminobenzidine (DAB) with a metal enhancer substrate for demonstrating quantitative differences in the amount of epidermal growth factor (EGF) in the submandibular gland from normal, castrated, and testosterone propionate (TP) treated castrated rats. Immunohistochemical determination of EGF visualized by DAB-nickel reagent was performed with image analysis using a computed image analyzer system (ACAS 570). Immunohistochemistry for EGF disclosed positive staining in granular convoluted tubule cells in the tissue sections from each experimental group. Using the tools of a competent data program installed in the ACAS 570 software, we measured quantitative differences among the experimental glands examined. Castration was shown to elicit a significant reduction in the EGF-positive area and staining intensity, and administration of TP to the castrated animals restored these parameters to levels greater than those of normal rats. Our study demonstrates that a simple, inexpensive, commercially available metal enhancer substrate can be applied accurately to the computer assisted quantification of histochemical hormone-induction studies.

3,3'-Diaminobenzidine↗
PubMed1979

[Methodical investigations for quantitative image analysis on cells of the female genital tract (author's transl)].

An earlier published [8] image analysis system for cytology is tested on gynecological material. At first the question for the staining method which is optimal for scanning-photometrical measurements is investigated. In addition the wavelength of the light which reproduces best the nuclear structure must be found. Feulgen-stained preparations scanned at 570nm allow a more secure sorting of superficial cells against intermediary cells than those in the classical Papanicolaou-stain at white light. Discoloring of Papanicolaou-preparations and Feulgen staining afterwards was accomplished without technical problems and there were not detectable any qualitative differences with directly Feulgen-stained preparations by means of image analysis. With this method it is possible to recognize structural changes in nuclei of vaginal epithelium during cell differentation.

Cell Nucleus↗