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Rapid detection of chlorine-induced bacterial injury by the direct viable count method using image analysis.

A modified direct viable count method to detect living bacteria was used with image analysis for the rapid enumeration of chlorine-injured cells in an Escherichia coli culture. The method was also used for determining chlorine-induced injury in coliform isolates and enteric pathogenic bacteria. Cultures were incubated in phosphate-buffered saline, containing 0.3% Casamino Acids (Difco Laboratories, Detroit, Mich.), 0.03% yeast extract, and optimal concentrations of nalidixic acid. Samples were withdrawn before and after incubation and stained with acridine orange, and cell lengths and breadths were measured by computerized image analysis. After incubation, cells which exceeded the mean preincubation length (viable cells) were enumerated and the results were compared with those obtained by the plate count method. Injury in the chlorine-exposed cell population was determined from the difference in viable count obtained with a nonselective Casamino Acids-yeast extract-nalidixic acid medium and a selective Casamino Acids-yeast extract-nalidixic acid medium containing sodium deoxycholate or sodium lauryl sulfate. The levels of injury determined by the direct viable count technique by using image analysis were comparable to those determined by the plate count method. The results showed that image analysis, under optimal conditions, enumerated significantly higher numbers of stressed E. coli than the plate count method did and detected injury in various cultures in 4 to 6 h.

Chlorine↗

The cumulative verification image analysis tool for offline evaluation of portal images.

PURPOSE: Daily portal images acquired using electronic portal imaging devices contain important information about the setup variation of the individual patient. The data can be used to evaluate the treatment and to derive correction for the individual patient. The large volume of images also require software tools for efficient analysis. This article describes the approach of cumulative verification image analysis (CVIA) specifically designed as an offline tool to extract quantitative information from daily portal images. METHODS AND MATERIALS: The user interface, image and graphics display, and algorithms of the CVIA tool have been implemented in ANSCI C using the X Window graphics standards. The tool consists of three major components: (a) definition of treatment geometry and anatomical information; (b) registration of portal images with a reference image to determine setup variation; and (c) quantitative analysis of all setup variation measurements. The CVIA tool is not automated. User interaction is required and preferred. Successful alignment of anatomies on portal images at present remains mostly dependent on clinical judgment. Predefined templates of block shapes and anatomies are used for image registration to enhance efficiency, taking advantage of the fact that much of the tool's operation is repeated in the analysis of daily portal images. RESULTS: The CVIA tool is portable and has been implemented on workstations with different operating systems. Analysis of 20 sequential daily portal images can be completed in less than 1 h. The temporal information is used to characterize setup variation in terms of its systematic, random and time-dependent components. The cumulative information is used to derive block overlap isofrequency distributions (BOIDs), which quantify the effective coverage of the prescribed treatment area throughout the course of treatment. Finally, a set of software utilities is available to facilitate feedback of the information for treatment plan recalculation and to test various decision strategies for treatment adjustment. CONCLUSIONS: The CVIA tool provides comprehensive analysis of daily images acquired with electronic portal imaging devices. Its offline approach allows characterization of the nature of setup variation for the individual patient that would have been difficult to deduce using only a few daily or weekly portal images. Distribution of the tool will help establish an important database of setup variation from many clinics. The information derived from CVIA can also serve as the foundation to integrate treatment verification, treatment planning, and treatment delivery.

Computer Peripherals↗

Applications of image analysis to anatomic pathology: realities and promises.

Image Analysis in Pathology is viewed as an ancillary method meant to provide objective support in the resolution of difficult problems. Its Achilles heel is the process of nuclear segmentation (delimitation of the nuclear membrane) which is extremely difficult in pathology materials. Although interactive segmentation procedures are available no reliable fully automatic method has been described. The only application of image analysis that has truly succeeded in Pathology is DNA ploidy measurement. A very desirable application is the quantitation of immunohistochemical markers, which is technically challenging, has been resolved only in certain cases and is unlikely to have a general solution. Nuclear quantitation has repeatedly proven to be helpful in reaching differential diagnoses, in particular when based on size distributions of nuclear profiles rather than its average, but is hampered by the segmentation problem discussed above. Texture analysis of chromatin is an exciting, mathematically complex application likely to succeed, for which many approaches have been described. Finally a diagnosis (classification) can be obtained based on algorithms applied to multiple descriptors of tumor cells (for instance nuclear sizes, chromatin texture, shape, etc). The best classificatory approaches are neural networks (a form of artificial intelligencee), multivariate analysis, and logistic regression (statistical).

Algorithms↗

[Computer-based image analysis for experimental and clinical morphology--principles, utilization and marginal limits].

The new computer based image analysis techniques are powerful tools for mophometrical and quantitative image analysis in case of clinical and experimental morphology. Digital image analysis requires a distinction between two phases 1. generation of fundamental data (x,y coordinates and grey values of the pixel) and 2. calculation of parameters from these data. Stereological procedures are very powerful in quantifying morphological phenomenons, but computer based image analysing techniques allow multiple analysis of morphological objects and analysis of statistical distributions. There is great scientific benefit using modern computer based image analysing techniques.

Animals↗

Phenotypic analysis of adults and eggs of Fasciola hepatica by computer image analysis system.

Knowledge of the morphological phenotypes of the liver fluke Fasciola hepatica (Trematoda: Digenea) is analysed. The influence of parasite age on its dimensions, the adult fluke growth model, variation in a biometric variable versus time, and variation in a biometric variable versus another biometric variable (allometric model) are revised. The most useful allometric model appears to be (y2 m-y2)/y2=c [(y1 m-y1)/y1](b), where y1=body area or body length, y2=one of the measurements analysed, y1 m, y2 m=maximum values towards which y1 and y2, respectively, tend, and c, b=constants. A method based on material standardization, the measurement proposal and allometric analysis is detailed. A computer image analysis system (CIAS), which includes a colour video-camera connected to a stereomicroscope (for adult studies) and a microscope (for egg studies), facilitates the processing of digital imaging. Examples of its application for the analysis of the influence of different factors on the liver fluke phenotype are shown using material from the Northern Bolivian Altiplano, where human and domestic animal fascioliasis is caused by F. hepatica only. Comparisons between the development of livestock fluke populations from highlands and lowlands are discussed and the relationships between host species and liver fluke morphometric patterns is analysed.

Altitude↗

Enamel demineralisation assessed by computerised image analysis of clinical photographs.

OBJECTIVES: To investigate the use of computerised image analysis to measure the area of demineralisation on the buccal surface of a tooth and to analyse the effect on that measurement of varying the angle at which a photographic image of the buccal surface of a tooth is taken. METHODS: Twenty-two teeth with areas of incremental demineralisation on the buccal surface were photographed perpendicular to the buccal surface and at angles of 20 and 40 degrees, above (Cuspal) and below (Gingival) the perpendicular. The images were captured and analysed using computerised image analysis. RESULTS: The Gingival views showed lower limits of agreement than the Cuspal views, probably due to the positioning of the masking on the lower part of the flash, to reduce reflection. There was a significant difference between the areas of demineralisation measured for the three angles from the Gingival views (p<0.001). The mean differences in areas measured were not linear, but there was a significantly greater reduction in area measured for the 40 degrees views. CONCLUSIONS: Computerised image analysis to measure areas of demineralisation from a captured 35mm slide is reproducible. The position of the masking on the camera flash and the angle of the camera makes a difference to the area measured and the grey scales recorded. Images taken at a larger angle than 20 degrees to the perpendicular will record a significantly reduced area of demineralisation. Standardisation of photographs with the use of calibration markers and manipulation of images is discussed.

Analysis of Variance↗

Use of image analysis techniques for objective quantification of the efficacy of different hair removal methods.

In the field of consumer-used cosmetics for hair removal and hair growth reduction, there is a need for improved quantitative methods to enable the evaluation of efficacy and claim support. Optimized study designs and investigated endpoints are lacking to compare the efficacy of standard methods, like shaving or plucking, with new methods and products, such as depilating instruments or hair-growth-reducing cosmetics. Non-invasive image analysis, using a high-performance microscope combined with an optimized image analysis tool, was investigated to assess hair growth. In one step, high-resolution macrophotographs of the legs of female volunteers after shaving and plucking with cold wax were compared to observe short-term hair regrowth. In a second step, images obtained after plucking with cold wax were taken over a long-term period to assess the time, after which depilated hairs reappeared on the skin surface. Using image analysis, parameters like hair length, hair width, and hair projection area were investigated. The projection area was found to be the parameter most independent of possible image artifacts such as irregularities in skin or low contrast due to hair color. Therefore, the hair projection area was the most appropriate parameter to determine the time of hair regrowth. This point of time is suitable to assess the efficacy of different hair removal methods or hair growth reduction treatments by comparing the endpoint after use of the hair removal method to be investigated to the endpoint after simple shaving. The closeness of hair removal and visible signs of skin irritation can be assessed as additional quantitative parameters from the same images. Discomfort and pain rating by the volunteers complete the set of parameters, which are required to benchmark a new hair removal method or hair-growth-reduction treatment. Image analysis combined with high-resolution imaging techniques is a powerful tool to objectively assess parameters like hair length, hair width, and projection area. To achieve reliable data and to reduce well known image-analysis artifacts, it was important to optimize the technical equipment for use on human skin and to improve image analysis by adaptation of the image-processing procedure to the different skin characteristics of individuals, like skin color, hair color, and skin structure.

Adult↗

[Digital image analysis in pathology--exemplified in prostatic cancer].

Digital image analysis is increasingly used in medicine. This paper reviews image analysis methods currently used in pathology. To exemplify established and future use of these methods, we present our own research and available literature on prostate cancer. DNA ploidy examinations mean measuring nuclear DNA content. A review of available literature shows that DNA ploidy is a very good prognostic marker in early and localized prostate cancer. Nevertheless, request for such examinations is sparse, in contrast to what is the case for gynecological cancers. In addition to assessing nuclear DNA content, we have today the means of quantitatively assessing the chromatin organisation. Such nucleotyping and texture analysis of chromatin has been shown to give significant diagnostic and prognostic information both in localized and advanced prostate cancer. DNA ploidy and nucleotyping are methods that analyze features of single cells. New methods for describing tissue architecture in an objective and reproducible way have been developed in our department. Preliminary results demonstrate the ability of these methods to discriminate between groups of prostate cancer patients with good or poor diagnosis. Digital techniques in combination with new methods from molecular biology have a potential for reducing workload, with parallel assessment of a large number of markers on large populations. The combination of traditional, molecular and digital pathology already offers the possibility of improved diagnostic and prognostic activity. However, there is the challenge of actually employing these methods. DNA ploidy for localized prostate cancer is only one example of a reliable prognostic marker that is not used to its full potential.

Chromatin↗

Digital pathology, image analysis, and artificial intelligence in liver disease.

Advances in digital pathology, image analysis, and artificial intelligence (AI) are rapidly transforming how pathologists and researchers interact with tissue samples and enable the development of diagnostic tools that harness high-resolution whole-slide images; these advances are in turn creating new opportunities for research, education, and routine clinical care globally. Liver disease is no exception, and digital pathology and AI have many applications in the diagnosis of liver cancer and liver diseases and in the assessment and management of transplantation. Although quantitative image analysis techniques have been applied to liver disease in research settings for over 50 years, recent improvements in image resolution, data storage, and the availability of advanced AI methods such as deep learning have driven multiple exciting developments. In this Review, we summarise the advancements in digital pathology, image analysis, and AI in liver disease. Key challenges such as access to and the logistics of using digital solutions, quality issues, and appropriate guidance in research and clinical use are reviewed, along with potential solutions to these challenges in the context of liver pathology and liver disease. Digital technologies are well established in liver pathology research, and access in clinical practice is increasing, with potential to address current laboratory challenges. Further evaluation is required to assess real-world effectiveness, clinical safety, and implementation of AI tools in liver pathology.

Journal Article↗

Ancillary techniques in the followup of transitional cell carcinoma: a comparison of cytology, histology and deoxyribonucleic acid image analysis cytometry in 91 patients.

PURPOSE: Voided urine and bladder washing cytology are used frequently in the evaluation of transitional cell carcinoma of the bladder. As part of an ongoing investigation we report on the role of deoxyribonucleic acid (DNA) image analysis cytometry as an adjunct to cytology in the followup of patients with transitional cell carcinoma. MATERIALS AND METHODS: Urine cytology and image analysis cytometry were performed independently on aliquots of voided urine, catheterized urine or bladder washings from 91 patients with previous or active transitional cell carcinoma of the bladder, and the results were compared to those of concurrent biopsy and clinical followup. RESULTS: Of 75 recurrent transitional cell carcinomas 42 were detected by cytology, while 63 and 64 were identified by image analysis cytometry and biopsy, respectively, for a sensitivity of 57, 84 and 85%, respectively. Combined cytology and image analysis cytometry detected 67 recurrences, for an overall sensitivity of 89%. Of 11 cases undetected by concurrent biopsy 9 had abnormal DNA histograms with transitional cell carcinoma at followup and 2 were DNA diploid but with grade 1 transitional cell carcinoma at followup. Of 12 cases undetected by image analysis cytometry 8 were grade 1 and 4 were grade 2 transitional cell carcinoma. CONCLUSIONS: Urine cytology and image analysis cytometry detect most recurrent tumors. Their combined use is indicated in the followup of patients with bladder transitional cell carcinoma.

Adult↗

Immunocytochemical assay for oestrogen receptor in fine needle aspirates of breast cancer by video image analysis.

Assessment of heterogeneity in oestrogen receptor (ER) expression aims to improve prediction of prognosis and treatment assignment in breast cancer. Current assessments are performed manually and are subjective. Automated image analysis as described here objectively quantitates ER in breast cancer nuclei obtained by needle aspiration. ER was visualised by ERICA with diaminobenzidine (DAB) substrate. Various indices of ER positivity were derived from the integrated density and average density measurements of nuclear DAB. Each index was compensated for background staining by non-specific antibody binding and endogenous peroxidase activity. Total nuclear ER content (integrated optical density of stain) was strongly associated with the biopsy ER concentration determined by saturation analysis of radioligand binding (DCC), P less than 0.005. Nuclear ER concentration by image analysis (mean optical density of stain) was not associated with the DCC measurement of ER concentration, P greater than 0.05. This was attributed to technical artefacts of cytocentrifugation. Using threshold values of 5% positive cells and 10 fmol mg-1 concordance of assignment of ER status by image analysis with the DCC assay was 91%, sensitivity was 89% and specificity 100%. It was concluded that image analysis is an appropriate, easy and economic method for determining the nuclear ER status of aspirated cancer cells. Image analysis has the potential to become a powerful diagnostic tool in the assessment of hormone receptor status of breast cancer patients.

Biopsy, Needle↗

An image analysis approach for automatically re-orienteering CT images for dental implants.

In the last decade, computerized tomography (CT) has become the most frequently used imaging modality to obtain a correct pre-operative implant planning. In this work, we present an image analysis and computer vision approach able to identify, from the reconstructed 3D data set, the optimal cutting plane specific to each implant to be planned, in order to obtain the best view of the implant site and to have correct measures. If the patient requires more implants, different cutting planes are automatically identified, and the axial and cross-sectional images can be re-oriented accordingly to each of them. In the paper, we describe the defined algorithms in order to recognize 3D markers (each one aligned with a missed tooth for which an implant has to be planned) in the 3D reconstructed space, and the results in processing real exams, in terms of effectiveness and precision and reproducibility of the measure.

Algorithms↗

Digital image analysis and stereology of angiogenesis in polypoid and nonpolypoid colorectal adenomas.

OBJECTIVE: To evaluate 3-dimensional parameters and bidimensional microvascular quantification in the different morphologic presentations of colorectal adenomas. STUDY DESIGN: A study was carried out, including 102 neoplastic colorectal lesions obtained by endoscopy or surgical resection. For the analysis of angiogenesis, immunohistochemistry, digital image analysis, microvascular quantification and stereology were used. RESULTS: Microvascular quantification, volume and microvascular length estimate rose gradually with high grade dysplasia as compared to the low grade ones (P < .001). There was no significant difference in angiogenesis between polypoid and nonpolypoid colorectal adenomas in terms of quantification and microvascular length estimate. CONCLUSION: The use of digital image analysis and stereology added greater objectivity and effectiveness to angiogenic evaluation because they allowed accurate segmentation of hypervascular areas, representation of the characteristic 3-dimensional morphology of the vascular supply and identification of differences in microvascularization in the developmental stages of colorectal cancer. However, no significant relation could be found between macroscopic type and angiogenesis, suggesting that angiogenesis may contribute little to morphogenesis of colorectal adenomas.

Adenoma↗

Measurement of anisotropic vertebral trabecular bone loss during aging by quantitative image analysis.

Age-dependent variations in the architecture of vertebral trabeculae in both the vertical and horizontal planes were characterized by quantitative image analysis. Images were obtained from autopsy specimens of the third lumbar vertebrae in 61 subjects (30 men and 31 women) whose ages ranged between 33 and 89 years). All subjects had died acutely either after trauma or illnesses unrelated to the skeleton. Using mathematical morphology techniques, we measured total bone area and perimeter, and the width of trabecular particles and medullary spaces in each slice. Between the age intervals 33-49 and 80-89 years: total bone loss in the vertical and horizontal planes was 51 and 64% for women, and 38 and 29% for men, respectively. Mean trabecular width (MTW) in the vertical plane decreased from 172 to 128 micron in women and from 181 to 144 micron in men; MTW in the horizontal plane fell from 144 to 112 micron in women and remained at 114 micron in men. Maximum trabecular width decreased with age in both planes in both sexes. The mode for trabecular width was 111 micron in both sexes for all ages and in both planes. The total number of trabeculae decreased only for women in the vertical plane. Intertrabecular spaces enlarged reciprocally as the trabeculae became thinner, but the widening of spaces was much greater than that expected with trabecular thinning alone. We conclude that age-related bone loss is comprised of two processes: reduction of MTW and fragmentation and complete loss of some trabeculae. We found no evidence of vertical trabeculae thickening during normal aging.

Adult↗

Assessment of bone mass by image analysis of metacarpal bone roentgenograms: a quantitative digital image processing (DIP) method.

a digital processing (DIP) method for assessing bone mass was developed on the basis of image analysis of roentgenograms. Linearity between DIP values and the actual calcium carbonate content was scarcely affected even if roentgenograms were made with bone phantoms placed in different depths of water or by altering the voltage of X-ray generation. In clinical studies, coefficients of variation (CV) for various measurements were lower than 2.4%. When the correlations between the DIP values and the bone mineral densities in the distal one-third of the radius, and the 2nd to 4th lumbar vertebrae were investigated in 340 females, there were good positive correlations of r = 0.799, and r = 0.611, respectively (p less than 0.001). The DIP value was significantly lower in patients showing a low Singh index and in those with vertebral fractures than in other subjects. These results suggest that the DIP method provides an index with which to assess the efficacy of treatment and which can be used as a criterion in screening for osteoporosis.

Absorptiometry, Photon↗

Image analysis assessment of the abnormal testis biopsy in male infertility.

PURPOSE: We previously demonstrated that testis biopsy image analysis is an effective method for quantifying intratubular spermatogenic cells in the obstructed testis with normal spermatogenesis. As an extension of the initial report, we describe using the quantitative ploidy and morphological characteristics of cells counted with image analysis in abnormal testis biopsies obtained for a male infertility evaluation. MATERIALS AND METHODS: Image analysis using a specifically designed filter was performed on Feulgen stained 5 microns, sections of paraffin embedded testicular tissue. Archival testicular tissue had been obtained using standard biopsy techniques from patients with azoospermia or severe oligospermia. Qualitative classification was based on standard evaluation of hematoxylin and eosin processed tissue. RESULTS: There were 62 biopsies performed in 58 men. Significant differences in the intratubular content of haploid (spermatozoa and spermatids), diploid and tetraploid cells were found among the 5 categories of abnormalities: the Sertoli-cell-only syndrome, spermatocyte arrest, spermatid arrest, hypospermatogenesis and normal spermatogenesis. Moderate variability was found in the proportion of cell types in spermatid arrest and hypospermatogenesis. CONCLUSIONS: Testis biopsy image analysis provides a quantitative method for categorizing abnormalities of intratubular cell content present in male infertility states by using deoxyribonucleic acid content and morphology characteristics. The limitations of the present qualitative analysis system are emphasized by the moderate variability evident within the current categories of spermatid arrest and hypo-spermatogenesis states.

Adult↗

Image analysis of radiolucencies in approximal enamel: a comparison of xeroradiographic and conventional film images.

Xeroradiographic and conventional film bitewing images of radiolucencies in approximal enamel were compared using a computer-aided image analysis system. Densitometric analysis of the conventional film types used in the study showed that the "lead foil free" film exhibited lower overall density when compared with conventional D-speed film. Computer-aided image analysis of radiolucencies in approximal enamel found that xeroradiographs depicted a higher degree of demineralization and a larger area than conventional radiography with the "lead foil free" film.

Adolescent↗

The automatic assessment of knee radiographs in osteoarthritis using digital image analysis.

We have developed an automatic system for the measurement of joint space in the knee using computerized analysis of digital stored images of knee radiographs. PA X-rays of the knee are positioned on a conventional viewing box. A video image of the radiograph is converted to digital form, stored and displayed on a closed circuit television monitor. This is edited and analysed by a microcomputer. After careful positioning under a computer generated grid on the television screen, automatic measurements are made using an edge detection facility. Two different edge detection algorithms were compared. Reproducibility is very good. Inter- and intra-observer relationships are also good with no significant difference between observers using a paired t-test and very good correlations. Results show that rapid, reliable and accurate measurement of joint space size can be achieved using digital image analysis. The application of image handling computers to radiological scoring is important in our understanding of joint destruction and the progression of the rheumatic diseases.

Algorithms↗