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Systematic investigations of the contrast results of histochemical stainings of neurons and glial cells in the human brain by means of image analysis.

The investigation of neurohistological specimens by image analysis has become an important tool in morphological neuroscience. The problems which arise during the processing of these images are non-trivial, especially if a pattern recognition of cells in the imaged tissue is intended. One of the major problems faced concerns the segmentation of structures of interest, whether cells or other histologic structures. The segmentation problem is often the result of an inappropriate staining procedure. For serious image analysis to be performed, the material under investigation must be optimally prepared. Spatially complex patterns, e.g. fuzzy-like neighbouring neurons, are easy to recognize for humans. But the integrative and associative performance of current artificial neuronal network schemes is too low to achieve the same recognition quality as humans do. Therefore, a general analysis of staining characteristics was performed, especially with respect to those stains which are relevant to object segmentation. Although most image analytical investigations of tissues are based on stained samples, a study of this type has not been previously conducted. Of the stains and procedures evaluated, the gallocyanin chrome alum combination staining provided the best stain contrast. Furthermore, this staining method shows sufficient constancy within different parts of the human brain. Even the fine nuclear textures are differentiable and can be used for further pattern recognition procedures.

Adult↗

Staining mast cells for morphometric evaluation on an image analysis system.

Multiple skin sections from three nonhuman primates (Macaca mulatta) and three hairless guinea pigs (Cavia porcellus) were stained with 12 different histologic stains to determine whether mast cells could be selectively stained for morphometric analysis using an image analysis system (IAS). Sections were first evaluated with routine light microscopy for mast cell granule staining and the intensity of background staining. Methylene blue-basic fuchsin and Unna's method for mast cells (polychrome methylene blue with differentiation in glycerin-ether) stained mast cell granules more intensely than background in both species. Toluidine blue-stained sections in the guinea pig yielded similar results. Staining of the nuclei of dermal connective tissue was enhanced with the methylene blue-basic fuchsin and toluidine blue stains. These two stains, along with the Unna's stain, were further evaluated on an IAS with and without various interference filters (400.5-700.5 nm wavelengths). In both the methylene blue-basic fuchsin and toluidine blue stained sections, mast cell granules and other cell nuclei were detected together by the IAS. The use of interference filters with these two stains did not distinguish mast cell granules from stained nuclei. Unna's stain was the best of the 12 stains evaluated because mast cell granule staining was strong and background staining was faint. This contrast was further enhanced by interference filters (500.5-539.5 nm) and allowed morphometric measurements of mast cells to be taken on the IAS without background interference.

Animals↗

Comparison of conventional morphometry and image analysis for the solution of histomorphometric problems.

The main differences between conventional morphometry and image analysis for their application in histopathology are pointed out. Conventional morphometry allows the use of contextural informations and a priori knowledge in a convenient manner. This is the most complicated thing in image analysis. The solution of this problem lies on the field of artificial intelligence. On the other hand, image analysis is able to measure densitometrical and textural features, to determine arbitrary quantiles and moments of feature distributions, to match scenes. Therefore the descriptive power of image analysis is much higher than the power of conventional morphometry.

Humans↗

Effects of lossy image compression on quantitative image analysis of cell nuclei.

OBJECTIVE: To investigate whether statistically significant changes occur in quantitative image analysis of cell nuclei when lossy image compression techniques are used. STUDY DESIGN: Thirty-five stoichiometric, Feulgen-stained samples of rat hepatocytes, human thyroid and ovarian cancer cell nuclei were used. Image analysis was performed by a computerized system that used AutoCyte LINK V1.1.1.56 software (Burlington, North Carolina, U.S.A.) for image acquisition and Zeiss Vision KS 400 V3.0 software (Oberkochen, Germany) for quantitative image analysis. After lossy JPEG compression of acquired images at different quality levels, some densitometric features were selected and measurements performed. RESULTS: We observed that nearly all the standard densitometric features showed statistically significant changes when images were compressed with the lossy JPEG algorithm. However, most invariant densitometric moment features remained free of statistically significant changes. CONCLUSION: The standard densitometric measurements that we used do not tolerate lossy compression. However, analyses using invariant densitometric features may be performed on images compressed with lossy JPEG, resulting in simpler, less expensive systems demanding less network bandwidth.

Animals↗

Automated Microarray Image Analysis Toolbox for MATLAB.

UNLABELLED: The Automated Microarray Image Analysis (AMIA) Toolbox for MATLAB is a flexible, open-source, microarray image analysis tool that allows the user to customize analyses of microarray image sets. This tool provides several methods to identify and quantify spot statistics, as well as extensive diagnostic statistics and images to evaluate data quality and array processing. The open, modular nature of AMIA provides access to implementation details and encourages modification and extension of AMIA's capabilities. AVAILABILITY: The AMIA Toolbox is freely available at http://www.pnl.gov/statistics/amia. The AMIA Toolbox requires MATLAB 6.5 (R13) (MathWorks, Inc. Natick, MA), as well as the Statistics Toolbox 4.1 and Image Processing Toolbox 4.1 for MATLAB or more recent versions. CONTACT: amanda.white@pnl.gov

Algorithms↗

[Analysis of the difference in NIH3T3 cell protein expression profiles before and after TPA treatment using two-dimensional polyacrylamide gel electrophoresis and image analysis software].

OBJECTIVE: To establish and optimize two-dimensional (2-D) polyacrylamide gel electrophoresis (PAGE) for comparative analysis of the protein expression profiles in mouse fibroblast NIH3T3 cells before and after 12-O-tetradecanoy- lphorbol-13-acetate (TPA) treatment using image analysis software, so as to prepare for more intensive study in the identification of the proteins that mediate the biological effect of TPA. METHODS: The total cellular protein extracted from NIH3T3 cells with or without TPA treatment underwent 2-D PAGE and silver nitrate staining prior to the analysis of the differential protein expressions using image analysis software. RESULTS: Image analysis revealed obvious differential protein expressions of the cells in response to TPA treatment. CONCLUSION: High-quality instruments for 2-D PAGE, skilled electrophoretic operation and efficient image processing are all essential in the reliable identification of stable functional proteins.

Animals↗

Assessment of hormone receptors in breast carcinoma by immunocytochemistry and image analysis. I. Progesterone receptors.

Frozen sections of 30 breast carcinomas were stained for progesterone receptors (PRs) using a rat monoclonal primary antibody and an alkaline phosphatase-antialkaline phosphatase technique. The micro-TICAS image analysis system was used for evaluation of the staining, with the results obtained by image analysis compared with the results of biochemical assays for PR. Strong positive/negative concordance (90%) was observed between the immunohistochemical and biochemical assays. However, the numerical values of the positive cases in the two assays did not correlate well, possibly because the biochemical assay does not take tumor cellularity into account. Three PR distribution patterns, designated A, B and C, were identified by image analysis among the breast tumors. In the type A pattern, tumor cell nuclei were diffusely and uniformly labeled. In type B, both clearly negative as well as distinctly positive cells were present. In type C tumors, a broad range of labeling reactions (from negative to intensely positive) was observed. These results imply (1) that the PR content of human breast carcinoma may be accurately and objectively assessed by the image analysis of immunohistochemically stained frozen sections, (2) that image analysis may provide a more accurate estimate of the cellular content of PR than do biochemical assays and (3) that PR distribution patterns obtained through image analysis permit the consistent appraisal of intratumoral heterogeneity of PR expression, which is potentially of prognostic importance.

Breast Neoplasms↗

Approach to diagnostic image analysis of melanocytic tumors.

Numerous attempts have been made to apply image analysis in dermatopathology. The technics used comprise measurement of nuclear size, shape, chromatin content, and texture, evaluation of immunohistological slides, assessment of proliferation, pattern analysis, and tumor volume estimation. For commonly accepted routine use, however, image analysis research has to be extended to large numbers of cases, using straightforward and reproducible measuring procedures, and to the development of ready-to-use equipment for specific tasks. In this way, image analysis in dermatopathology might supply useful diagnostic tools in addition to conventional microscopy, and may increase our understanding of morphology as a whole.

Diagnosis, Differential↗

Quantitation of histochemical staining of salivary gland mucin using image analysis in cats and dogs.

Two different, computer-based, image analysis methods were employed to complement subjective assessment of patterns of mucin staining (by periodic acid Schiff/alcian blue, aldehyde fuchsin/alcian blue and potassium hydroxide-alcian blue/phenylhydrazine hydrocholoride) in digitised images of sections of major and minor salivary glands from cats and dogs. Image analysis based on red, green and blue (RGB) staining was not suitable for quantitation of histochemical staining of mucin in salivary glands. Image analysis based on hue, saturation and lightness (HSL) allowed quantitative assessment of staining by different stain components and of mixed staining, and enabled comparison of staining of different glands in dogs and cats. Quantitative analysis based on HSL allowed differentiation of differences in staining patterns of major and minor cat and dog salivary glands, and between the species; such differences would have been impossible to distinguish on subjective grounds alone. Quantitative assessment of normal salivary gland histochemistry allows comparison with staining patterns in disease.

Animals↗

[Quantitative evaluation of nystagmus by an image-analysis system].

PURPOSE: We attempted to apply a newly developed image-analysis system for measurement and analysis of nystagmus. METHOD: Eye movements were recorded by digital video through a head-mounted charge coupled device (CCD) camera. The recorded movie was converted into black and white in order to detect the area of the pupil. Horizontal and vertical eye positions were determined by calculating the centroid of the pupil. Torsional angle was calculated using the iris striate pattern around the pupillary margin. RESULTS: The parameters (amplitude, cycle, etc.) of nystagmus were calculated easily by the new image-analysis system from the recorded images. As examples, the foveation period was measured accurately in a case of jerky-type congenital nystagmus. Very regular cycles of intorsional attack period were revealed in a case of superior oblique myokymia. A case of cork-screw-like nystagmus showed a characteristic combination of large and small cycles unassociated with torsion. CONCLUSION: This image-analysis system was useful for quantitative analysis of nystagmus, and especially for measurement of torsion. Detailed waveforms and specific rhythms of nystagmus, which could not be recognized by observation, were demonstrated by this system.

Adult↗

Multiple sclerosis medical image analysis and information management.

Magnetic resonance imaging (MRI) has become a central tool for patient management, as well as research, in multiple sclerosis (MS). Measurements of disease burden and activity derived from MRI through quantitative image analysis techniques are increasingly being used. There are many complexities and challenges in building computerized processing pipelines to ensure efficiency, reproducibility, and quality control for MRI scans from MS patients. Such paradigms require advanced image processing and analysis technologies, as well as integrated database management systems to ensure the most utility for clinical and research purposes. This article reviews pipelines available for quantitative clinical MRI research in MS, including image segmentation, registration, time-series analysis, performance validation, visualization techniques, and advanced medical imaging software packages. To address the complex demands of the sequential processes, the authors developed a workflow management system that uses a centralized database and distributed computing system for image processing and analysis. The implementation of their system includes a web-form-based Oracle database application for information management and event dispatching, and multiple modules for image processing and analysis. The seamless integration of processing pipelines with the database makes it more efficient for users to navigate complex, multistep analysis protocols, reduces the user's learning curve, reduces the time needed for combining and activating different computing modules, and allows for close monitoring for quality-control purposes. The authors' system can be extended to general applications in clinical trials and to routine processing for image-based clinical research.

Humans↗

Quantitative Evaluation of Nystagmus by an Image-analysis System.

Purpose: We attempted to apply a newly developed image-analysis system for measurement and analysis of nystagmus.Method: Eye movements were recorded by digital video through a head-mounted charge coupled device (CCD) camera. The recorded movie was converted into black and white in order to detect the area of the pupil. Horizontal and vertical eye positions were determined by calculating the centroid of the pupil. Torsional angle was calculated using the iris striate pattern around the pupillary margin.Results: The parameters (amplitude, cycle, etc.) of nystagmus were calculated easily by the new image-analysis system from the recorded images. As examples, the foveation period was measured accurately in a case of jerky-type congenital nystagmus. Very regular cycles of intorsional attack period were revealed in a case of superior oblique myokymia. A case of cork-screw-like nystagmus showed a characteristic combination of large and small cycles unassociated with torsion.Conclusion: This image-analysis system was useful for quantitative analysis of nystagmus, and especially for measurement of torsion. Detailed waveforms and specific rhythms of nystagmus, which could not be recognized by observation, were demonstrated by this system.

Journal Article↗

Osteometry by computer-aided image analysis: application to the human atlas.

Computer-assisted image-analysis having almost not been applied to macroscopical anatomy, particularly to osteometry, we used it for the automatic measurement of 8 osteological parameters on a series of 150 human atlases. From these measured parameters, 5 parameters have been directly calculated. The values obtained by image analysis and by measurement with vernier calliper are identical and similar to the data of the literature. The accuracy, the sources of error, and the great advantages of the image analysis method are then discussed.

Calibration↗

Automated image analysis to improve bead ingestion toxicity test counts in the protozoan Tetrahymena pyriformis.

AIMS: To improve bead ingestion counts in Tetrahymena pyriformis by automated image analysis as an alternative to direct-counts. METHODS AND RESULTS: Fluorescent latex beads were added to T. pyriformis cultures for ingestion tests. The number of beads ingested by 25 cells was counted directly by epifluorescence microscopy and compared with similar data from image analysis. anova indicated that counts were not significantly different (P < 0.05). The image analysis particularly provided advantages in terms of speed. CONCLUSIONS: The image analysis is superior to direct beads counting in T. pyriformis particularly in terms of speed of analysis. SIGNIFICANCE AND IMPACT OF THE STUDY: The image analysis method is very rapid and will allow many more toxicological analyses to be undertaken with less operator error.

Animals↗

One year's experience with two different image analysis systems for automated reading of the contrast fluorescence test.

We have tested two different personal-computer-based color image analysis systems for automated reading of the microlymphocytotoxicity test (LCT) for HLA-A,B,C typing and screening. Over 17,000 single LCT reactions were prepared using the simultaneous double fluorescent variant of the LCT (contrast fluorescence test, CFT). All tests were read visually by experienced laboratory staff members. For digital image analysis, an automated scanning system was used. The reactions were first recorded on a videotape recorder using a color (CCD) videocamera und subsequently analyzed with the two different image analysis systems by specifically developed programs. Good correlation (r = 0.89) of the score values assigned by digital image analysis with the visual tray reading was obtained. Since also the other main performance characteristics of the prototype system were acceptable for routine application, we may conclude that digital image analysis is a feasible and very interesting new technique for automated evaluation of the LCT.

Algorithms↗

[Electronic image analysis in ophthalmology].

Based on experimental investigations a broad concept for the use of television image analysis in basic and clinical ophthalmology is given. This summary outline contents a short description of the technique and some examples for the clinical use of image analysis, like the measurement of corneal width and erosions, infrared pupillography, morphometry of the iris and quantitative fluorescence angiography. "Static" and "dynamic" image analysis are defined, the role of pattern recognition is mentioned. The results of the study demonstrate that television image analysis can be of great importance for the future of quantitative ophthalmology.

Corneal Diseases↗

One year's experiences with two different image analysis systems for automated reading of the contrast fluorescence test.

We have tested two different personal-computer based color image analysis systems for automated reading of the microlymphocytotoxicity test (LCT) for HLA-A,B,C-typing and screening. Over 17,000 single LCT-reactions were prepared using the simultaneous double fluorescent variant of the LCT (contrast fluorescence test, CFT). All tests were read visually by experienced laboratory staff members. For image analysis, an automated scanning system was used. In a first step, reactions were recorded on a videotape recorder using a color(CCD)-video camera. In a second step, the recorded reactions were analyzed with the two different image analysis systems by specifically developed programs. Good correlation (r = 0.89) of the score values assigned by digital image analysis with the visual tray reading was obtained. Since also the other main performance characteristics of the prototype system (throughput, reliability, compatibility) were acceptable for routine application, we may conclude that digital image analysis is a feasible and very interesting new technique for automated evaluation of the LCT.

Algorithms↗

Evaluation of a new slide-based laser scanning cytometer for DNA analysis of tumors. Comparison with flow cytometry and image analysis.

DNA measurements generated by a new automated slide-based cytometer, the laser scanning cytometer (LSC), were compared with those produced by commercial flow cytometry (FCM) and image analysis (IA) devices. Laser scanning-cytometric analysis was performed by scanning alcohol-fixed, propidium iodide-stained tumor imprints with a 5-microns spot laser beam. Fifty-three malignant tumors (51 breast carcinomas and 2 lung carcinomas) were studied. Ploidy concordance rates for FCM versus LSC, IA versus LSC, and FCM versus IA were 96%, 91%, and 91%, respectively. Statistically significant agreement between methods was determined by linear regression analysis of DNA indices. Synthesis-phase fractions generated by FCM and LSC also were comparable, as demonstrated by linear regression (r = .83). Mean coefficients of variation for the LSC compared favorably with those for FCM and IA. The few discrepancies in ploidy status between methods could be explained by sampling error, the presence of possible near-diploid aneuploid populations that could not be effectively resolved by one or another modality, and the visual selection bias with IA when small aneuploid cell populations were present. The LSC shares many useful features with FCM, including automation, accuracy of quantitation, rapidity, and generation of reliable information regarding cell proliferation (synthesis-phase fraction). In addition, it has some of the advantages of IA, such as minimal tissue requirement, no need for special preparation, and the potential for visual selection of the cells measured. The LSC holds great promise for use in the clinical laboratory because of these combined characteristics.

Breast Neoplasms↗