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Detecting differences in crown transparency assessments between countries using the image analysis system CROCO.

Estimates of crown transparency calculated using the semi-automatic image analysis system CROCO were compared with the visual estimates of survey teams from 12 European countries. For each of five European species (Norway spruce (Picea abies (L.) Karst.), silver fir (Abies alba Mill.), Scots pine (Pinus sylvestris L.), beech (Fagus sylvatica L.), oak (Quercus robur L.)) 15 trees were assessed. In 64% of the comparisons, CROCO estimates and the survey teams' scores differed significantly, while differences between countries were significant in 58% of all comparisons. We calculated Spearman's rank correlation and the standard deviation of the differences between CROCO and each country and between pairs of countries. We then computed the medians for CROCO and each country. CROCO and only a few countries had for all species a higher median correlation and lower median standard deviation than the median for the comparison between countries. In addition no country had a consistently higher correlation or lower standard deviation than CROCO. We conclude that CROCO can provide more consistent and less variable estimates of crown transparency than visual assessments and that it can serve as a reference to detect differences in visual transparency assessments between countries over time.

Air Pollutants↗

Quantification of human sperm morphology and motility by means of semi-automatic image analysis systems.

Sperm morphology and motility are determined using a semi-automatic image analysis system under opto-manual control. The system consists of a microscope equipped with a drawing-tube, a digitizer-tablet, and a cursor interfaced to a small microcomputer. A light-emitting diode is mounted on the cursor, visible as a bright red pilot-light in the microscopic field. Sperm head and midpiece as well as the pathway of motile spermatozoa are traced with the cursor's red pilot light. The microcomputer calculates area, circumference, and length simultaneously. Motility and sperm density are determined altogether using microchamber technique. In a selected group of male subjects with normal, doubtful, and pathological semen analyses biometrical analyses of sperm morphology and motility were performed. Sperm morphology is best described by the variation of the head size and by the average midpiece width. Motility is best described by % motile spermatozoa and mean velocity determined 1 hour after semen collection. Biometrical semen analyses are superior to subjective evaluations regarding degree of information, objectivity, and reproducibility.

Cell Survival↗

Quantification and classification of human sperm morphology by computer-assisted image analysis.

A quantitative, semi-automated method for classifying human sperm based on objective measurements of head shapes and sizes has been developed. Air-dried smears of semen from eight healthy men were stained with the Feulgen reaction and 283 sperm were selected as prototypic examples of the 10 morphology classes used in our classification system. Sperm heads were imaged through a microscope (NA = 1.3), sampled at 0.125-micron intervals, and measured on an image analysis system. Measurements included stain content, length, width, perimeter, area, and arithmetically derived combinations. Additionally, each sperm image was optically sectioned at right angles to its major axis to give a measure of lengthwise heterogeneity of shape. Linear stepwise discriminant analysis was used to identify the more powerful parameters and to create a model employing eight parameters. The jackknifed classification procedure distinguished normal from abnormal sperm with 95% accuracy and correctly assigned 86% of the sperm to one of 10 shape classes. Most of the misclassification errors occurred among closely related classes. The results demonstrate the ability of automated image analysis to classify individual sperm into clinically familiar shape categories.

Humans↗

Breast imaging with fluorine-18-FDG PET: quantitative image analysis.

UNLABELLED: This study evaluated various quantitative criteria for analysis of breast imaging with PET using the radiolabeled glucose analog 18F-fluorodeoxyglucose (FDG). METHODS: In a prospective study, 73 patients with abnormal mammography or palpable breast masses scheduled for biopsy were investigated with PET. A total of 97 breast tumors were evaluated by histology, including 46 benign and 51 malignant tumors. Using a whole-body PET scanner, attenuation-corrected images were acquired between 40 and 60 min after tracer injection. For Patlak analysis, dynamic data acquisition was obtained in 24 patients. To differentiate between benign and malignant breast tumors, receiver operating characteristic curves were calculated using incrementally increasing threshold values for tumor/ nontumor ratios based on average and maximum activity values per region of interest, standardized uptake values (corrected for partial volume effect, normalized to blood glucose, partial volume effect and blood glucose, using the lean body mass as well as the body surface area) and calculating the FDG influx rate (K) assessed by Patlak analysis. RESULTS: Quantification of FDG uptake in breast tumors provided objective criteria for differentiation between benign and malignant tissue with similar diagnostic accuracy as compared with visual analysis. Applying correction for partial volume effect and normalization by blood glucose yielded the highest diagnostic accuracy. CONCLUSIONS: These quantitative methods provided accurate evaluation of PET data for differentiating benign from malignant breast tumors. Quantitative assessment is recommended to complement visual image interpretation with the potential benefit of reduced interobserver variability.

Breast Diseases↗

[Marketing orientation in rehabilitation--exemplified by image analysis of a rehabilitation clinic for addictions].

This article presents the findings of an image analysis carried out for a rehabilitation clinic for addictive diseases. The cross-sectional study was designed as an anonymous survey sent to relevant (regional) referral agencies (n = 90), e.g. counselling services for addictive diseases. The study covers the clinic's actual and ideal image, discriminating frequency of referrals, diagnostic categories, region, and type of institution. Results concerning the real profile indicate that on average the dimensions cooperation, communication with regard to personal contact, and reputation of the clinic were more critically judged than the issues information, communication with regard to discharge reports, standards of quality, and consideration of patients' needs. Cooperation, communication with regard to personal contact, and quality standards turned out to be the most important criteria from the referring agencies' perspective (ideal profile). A statistical comparison between those with high and low assignment rates suggested that customers rated the clinic more favourably than non-customers. Thus, it seems that the clinic's image has considerable influence on referral decisions. Particularly interesting is the additional finding that the referring institutions perceive pension insurers' directives to be of secondary relevance.

Alcoholism↗

Stereotactic investigation of limbic epilepsy using a multimodal image analysis system. Technical note.

A methodology has been developed for stereotactic investigation of limbic epilepsy using an image-analysis system that simultaneously displays different structural and functional images of the brain. The validity and accuracy of this system were established with phantom studies. Surgical planning and electrode implantation are guided by stereotactic magnetic resonance imaging, digital subtraction angiography, and position emission tomography. This methodology provides the spatiotemporal relationship of cerebral structure and function necessary to identify seizure onset and propagation in human limbic system epilepsy.

Brain Mapping↗

Effect of colchicine on atherosclerosis. III. Study of dermal elastic fibers by quantitative histochemistry, automated image analysis.

Computerized automatic-image analytical procedure was applied on dermal biopsies stained for elastin by a new procedure giving a completely white background and staining only the elastic fiber system. In arteriosclerotic hypertensive patients, a 3-4 months' treatment with 1 mg colchicine per day resulted in a significant (60-80%, p less than 0.01) increase of the dermal elastic fiber density both in the superficial papillary dermis and in the deep dermis. This result shows that the age-dependent increase of elastic fibers can be influenced by pharmacological means. The inhibition by colchicine of the synthesis and secretion of the fibroblast-derived metalloelastase-type protease could be a plausible explanation of this finding.

Adult↗

Semiautomatic image analysis for grain counting in in situ hybridization experiments.

We have developed a computer image analysis procedure for counting autoradiographic grains in in situ hybridization experiments. The procedure automatically estimates the number of autoradiographic grains over cells and measures cell number and size so that grain density per unit cell area can be calculated. Advantages include the clear separation of grains and cells, using chromatic and spatial filters to enhance the image; the use of gray level operators to extract cells from grains; and the use of binary operators for separating apposed or partially overlapping cells and grains. Comparison of manual and automated grain counts revealed a significant correlation between human and computer estimations of grain number. However, the automatic grain counting technique consistently underestimated the number of grains when grain density was high. Measures of the fractional area occupied by grains normalized by the average area of a single grain were a better estimate at high grain densities. The procedure can be modified easily to operate on most image analyzers.

Animals↗

Light microscopy and image analysis of thin filament lengths utilizing dual probes on beef, chicken, and rabbit myofibrils.

Image analysis procedures for immunofluorescence microscopy were developed to measure muscle thin filament lengths of beef, rabbit, and chicken myofibrils. Strips of beef cutaneous trunci, rectus abdominis, psoas, and masseter; chicken pectoralis; and rabbit psoas muscles were excised 5 to 30 min postmortem. Fluorescein phalloidin and rhodamine myosin subfragment-1 (S1) were used to probe the myofibril structure. Digital images were recorded with a cooled charge-coupled device controlled with IPLab Spectrum software (Signal Analytics Corp.) on a Macintosh operating system. The camera was attached to an inverted microscope, using both the phase-contrast and fluorescence illumination modes. Unfixed myofibrils incubated with fluorescein phalloidin showed fluorescence primarily at the Z-line and the tips of the thin filaments in the overlap region. Images were processed using IPLab and the National Institutes of Health's Image software. A region of interest was selected and scaled by a factor of 18.18, which enlarged the image from 11 pixels/microm to approximately 200 pixels/microm. An X-Y plot was exported to Spectrum 1.1 (Academic Software Development Group), where the signal was processed with a second derivative routine, so a cursor function could be used to measure length. Fixation before phalloidin incubation resulted in greatest intensity at the Z lines but a more-uniform staining over the remainder of the thin filament zone. High-resolution image capture and processing showed that thin filament lengths were significantly different (P < 0.01) among beef, rabbit, and chicken, with lengths of 1.28 to 1.32 microm, 1.16 microm, and 1.05 microm, respectively. Measurements using the S1 signal confirmed the phalloidin results. Fluorescent probes may be useful to study sarcomere structure and help explain species and muscle differences in meat texture.

Actin Cytoskeleton↗

Quantitative image analysis in mammary gland biology.

In this paper we present a summary of recent quantitative approaches used for the analysis of macro and microscopic images in mammary gland biology. The advantages and disadvantages of whole mount analysis, reconstruction of serial tissue sections and nucleus/cell segmentation of either conventional and confocal images are discussed, as are applications of quantitative image analysis, such as quantification of protein levels or vasculature measurements in normal tissue and cancer. Integration of quantitative imaging into the further study of the mammary gland holds the promise of better understanding its tissue complexity that evolves during development, differentiation and disease.

Animals↗

A computerized image analysis system for estimating Tanner-Whitehouse 2 bone age.

A method for assigning Tanner-Whitehouse 2 skeletal maturity scores (or bone ages) to hand-wrist X-rays by an image analysis computer system is described. An operator positions the relevant area of the X-ray on a light box beneath a video camera. Correct positioning is assured by computer templates of each bone stage. Thereafter the process is automatic; the computer, not the operator, rates the bones. The system produces continuous stage scores, not discrete ones such as B, C or D. Data are given which show that the computer-assisted skeletal age score is more repeatable than the usual manual (or unassisted) rating. The absolute difference between duplicates averaged 0.25 stages; differences of as much as 1.0 stage occurred in only 3% of duplicates compared with 15% obtained in manual ratings.

Age Determination by Skeleton↗

Automatic on-line electronic portal image analysis with a wavelet-based edge detector.

A fully automatic method for on-line electronic portal image analysis is proposed. The method uses multiscale edge detection with wavelets for both the field outline and the anatomical structures. An algorithm to extract and combine the information from different scales has been developed. The edges from the portal image are aligned with the edges from the reference image using chamfer matching. The reference is the first portal image of each treatment. The matching is applied first to the field and subsequently to the anatomy. The setup deviations are quantified as the displacement of the anatomical structures relative to the radiation beam boundaries. The performance of the algorithm was investigated for portal images with different contrast and noise level. The automatic analysis was used first to detect simulated displacements. Then the automatic procedure was tested on anterior-posterior and lateral portal images of a pelvic phantom. In both sets of tests the differences between the measured and the actual shifts were used to quantify the performance. Finally we applied the automatic procedure to clinical images of pelvic and lung regions. The output of the procedure was compared with the results of a manual match performed by a trained operator. The errors for the phantom tests were small: average standard deviation of 0.39 mm and 0.26 degrees and absolute mean error of 0.31 mm and 0.2 degrees were obtained. In the clinical cases average standard deviations of 1.32 mm and 0.6 degrees were found. The average absolute mean errors were 1.09 mm and 0.39 degrees. Failures were registered in 2% of the phantom tests and in 3% of the clinical cases. The algorithm execution is approximately 5 s on a 168 MHz Sun Ultra 2 workstation. The automatic analysis tool is considered to be a very useful tool for on-line setup corrections.

Automation↗

Ploidy of smooth muscle tumours: retrospective image analysis study of formalin fixed, paraffin wax embedded tissue.

The feasibility of using an image analyser, the CAS 100, to assess the ploidy of smooth muscle tumours on formalin fixed paraffin wax embedded tissue was assessed. Various different methods of assessment were compared. A paraffin wax block of 15 leiomyomata and 13 leiomyosarcomata yielded serial sections of 3 microns, 5 microns, and 7 microns and a cytospin preparation (from a 50 micron section). Sections and cytospin preparations were stained with Feulgen and quantified using the CAS 100. The suitability of lymphocytes, previously suggested to be unsuitable as control cells in tissue sections due to the compact nature of their DNA, was assessed in sections and cytospin preparations and compared with endothelial cells, the standard alternative, on the same slide. Despite having a mean nuclear area of only 18.5 microns 2 in sections lymphocytes had a similar diploid peak--that is, 4.1 pg--to endothelial cells (mean nuclear area 39 microns 2). A comparison of 3 microns, 5 microns, and 7 microns sections showed 5 microns to be the optimal thickness. Cytospin preparations yielded histograms of superior quality than those from tissue sections. All 15 leiomyomata had a 5C exceeding rate of less than 0.3%. Ten of 11 histologically malignant tumours had a 5C exceeding rate of more than 5% (mean 14%) in sections and cytospins. Two leiomyosarcomata that had arisen in the lower gastrointestinal tract had a mitotic rate of less than or equal to 1/10 per high power fields and yielded histograms similar to those of the leiomyomata. It is concluded that formalin fixed, paraffin wax embedded tissue can be used for DNA quantification by image analysis; that tissue sections yield poorer results than cytospin preparations; that lymphocytes are reliable control cells in cytospin preparations; and that a population of cell greater than 5C is seen in 90% of leiomyosarcomata.

Cell Separation↗

[Computerized image analysis of normal optic discs].

A self-designed computer system was used for image analysis of 172 normal optic discs, with Littmann's correction of photographic magnification. The mean disc area was found to be 2.40 +/- 0.50mm2, and the mean rim area 1.77 +/- 0.32mm2. The rim area was significantly correlated to disc area (r = 0.67, P = 0), but not to age, sex or refraction.

Adolescent↗

Characterization of chromatin structure by image analysis--a method for the assessment of changes in chromatin organization.

This paper describes an image analysis technique for the assessment of changes in chromatin organization in ultrathin tissue sections. Transmission electron micrographs of tissue sections were analyzed by a SEM-IPS image processing unit (Kontron) at a total magnification of 7,500. The boundaries of each nucleus and each nucleolus were defined interactively, and the grey level threshold between heterochromatin and euchromatin was determined. The grey level distribution and the total area of each nucleus was measured. In addition, the total area, number, and individual areas of heterochromatin particles and nuclei were measured. Based on these measurements, a number of different variables have been defined, and several of these have proved to discriminate between normal and malignant cells.

Animals↗