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PubMed2001

Application of computerized image analysis in pigmentary skin diseases.

BACKGROUND: Melanocyte number and the amount of melanin pigment are related to diagnosis and treatment of pigmentary skin diseases. Various histologic methods are used, such as Fontana-Masson stain for melanin pigment or immunohistochemical stain for melanocytes. Recently, computerized image analysis has been applied to many fields to avoid interobserver bias. In this study, we applied a computerized image analysis to assess the melanin content and melanocyte density of human epidermis. METHODS: We evaluated the skin biopsy specimens (paraffin blocks) from normal human skin (33 +/- 6.6, n = 11) and diseased skins; vitiligo (32 +/- 10.0, n = 8), melasma (35 +/- 8.6, n = 11), and lentigo senilis (40 +/- 7.2, n = 11) (mean age +/- SD). Each specimen was stained with Fontana-Masson for melanin pigments and immunohistochemical method for melanocytes. Quantitative analysis of melanin pigment and melanocyte number (density) were investigated through two methods: (1) two dermatologists measured the visual scales; and (2) computerized image analysis was used to measure melanin content indices (MCI). The data were evaluated using one-way ANOVA. RESULTS: The visual scale of the Fontana-Masson stain was the highest for lentigo senilis (3.8 +/- 0.40), followed by melasma (2.6 +/- 0.67), normal skin (1.8 +/- 0.60) and vitiligo (0) (P < 0.05). These findings were consistent with objective measurements made by computerized image analysis. MCI values were 120.3 +/- 20.74 for lentigo senilis, 81.1 +/- 19.27 for melasma, 45.5 +/- 16.92 for normal skin, and 0.3 +/- 0.30 for vitiligo in decreasing order (P < 0.05). MC/1E (melanocyte number per 1 mm epidermis) was about two fold larger in lentigo senilis (18.1 +/- 8.92) than melasma (9.7 +/- 2.40) or normal skin (9.3 +/- 2.67) (P < 0.05). MC/1B (melanocyte number per 1 mm basal layer) was about 1.5 fold higher in lentigo senilis (13.5 +/- 4.17), compared to normal skin (9.0 +/- 3.55) (P < 0.05). Melasma showed increased melanocyte numbers compared to normal skin, but it was not statistically significant (P > 0.05). CONCLUSION: We believe this computerized image analysis could be useful tool for diagnosis and comparison of interval changes in pigmentary diseases like melasma or lentigo senilis by quantifying melanin pigments or melanocytes in skin biopsy specimens.

Biopsy↗
PubMed1994

Use of image analysis to measure handwashing effectiveness.

BACKGROUND: This study tested a unique quantification technology (image analysis of fluorescent particles) for the measurement of handwashing effectiveness, comparing an automated system with traditional manual handwashing. In this instance, handwashing "effectiveness" was defined as ability of the technique to remove surface soil, as opposed to removal of microorganisms. METHODS: Sixty-five subjects washed their hands by manual handwashing (day 1) and automated handwashing (day 2). Hands were coated with an oil-based fluorescent suspension, photographed under controlled conditions, washed for 10 seconds, and rephotographed. Photographs from before and after handwashing were subjected to image analysis. RESULTS: Manual handwashing removed 73.1% of the particles; the automated method removed 23.2%. Parameters for use of image analysis as a technique to examine removal of gross contamination from the hands are given in the text. CONCLUSIONS: Although this method will not replace traditional culture methods for examining microbial removal, it is useful for measuring removal of particles (gross contamination) from skin or other surfaces. Under the conditions tested, manual handwashing was found to be decidedly superior to use of an automated system for removal of oil-based soil on the hands.

Automation↗
PubMed2000

Analysis of film coating thickness and surface area of pharmaceutical pellets using fluorescence microscopy and image analysis.

A method is presented which enables geometrical characterisation of pharmaceutical pellets and their film coating. It provides a high level of details on the single pellet level. Image analysis was used to determine the coating thickness (h) applied on the pellets and the surface area (A) of the pellet cores. Different definitions of A and h are evaluated. Hierarchical analysis of variance was used to resolve different sources contributing to the total variance. The variance within pellets and the variance between pellets were found as significant sources of variation. Special emphasis was put on evaluation of A/h due to its influence on the release rate of an active drug substance from the pellet core. The pellet images were thus used to predict variations in the release rate using a mathematical model as a link between the image data and the release rate. General aspects of image analysis are discussed. The method would be useful in calibration of near infrared spectra to h in process analytical chemistry.

Microscopy, Fluorescence↗
PubMed2000

Novel image analysis of corpus cavernous tissue in impotent men.

OBJECTIVES: To objectively evaluate the contents of corpus cavernous tissue in impotent men using an automated novel image analysis system. METHODS: Thirty-three impotent men and 2 normal potent men (controls) underwent corpus cavernous biopsies. The procedures were performed using a Biopty gun under local anesthesia. The obtained specimens were stained with Masson's trichome technique, and the collagen fiber contents were evaluated by a computerized morphometric analysis method. In addition, we estimated the intraobserver and interobserver reliability of this automated image analysis system. RESULTS: No major complication was noted during or after the biopsies. Of the 33 impotent patients, 3 were diagnosed as having psychogenic, 11 as having arteriogenic, 13 as having venogenic, 1 as having neurogenic, and 5 as having idiopathic impotence. The collagen fiber percentages in the 35 men were as follows: normal potent: 48.2% +/- 1.4%, psychogenic 55.2% +/- 11.6%, arteriogenic 73.2% +/- 4.4%, venogenic 66.5% +/- 4.2%, neurogenic 76.9%, and idiopathic 77.4% +/- 4.2%. Significant differences were found between the normal potent and arteriogenic groups (P <0.05) and between the normal potent and idiopathic groups (P <0.05). Patients older than 60 years had a higher collagen fiber content (70.4% +/- 3.7%) than those younger than 50 years old (58.6% +/- 5.2%). The interobserver and intraobserver variances were both negligible for this automated image analysis system. This method significantly reduced the amount of variation introduced by the intra-rater reliability of a technician compared with the manual method. CONCLUSIONS: The present automated image analysis system is believed to be a reliable, accurate quantitative measurement tool for studies of penile tissue. Cavernous biopsy is a rapid, safe, and representative modality to study penile disease. An increase in cavernous collagen fibers (or corporal fibrosis) is considered an important factor in impaired erectile function.

Adult↗
PubMed2004

Computer image analysis for measuring lean and fatty areas in cross-sectioned dry-cured hams.

The aims of this study were 1) to apply computer image analysis to obtain measures of lean and fatty areas on the cross section of dry-cured hams, 2) to investigate variation of these measures, and 3) to evaluate reproducibility and repeatability of these techniques. Traits of concern were the cross-sectional area (SA), lean, or muscles, area (LA), and the fatty area (FA) centered on the cross section and surrounded by biceps femoris, semimembranosus, semitendinosus, and quadriceps femoris, as well as the FA-to-SA ratio (FESR). Hams were obtained from crossbred pigs (n = 279) slaughtered at 9 mo of age (mean BW of 169 +/- 17 kg). Digital images of the cross section of dry-cured hams were captured using standardized procedures. Three replicated measures of areas were collected by three operators using three image analysis techniques (automatic, automatic-assisted, and manual). Variance components were estimated using a linear model that included slaughter group, gender, and gender x slaughter group as fixed effects and operators, pig, and operator x pig as random effects. Statistical analyses considered all measures (n = 7,533) or measures collected after reinstruction of all operators for spatial calibration of the analysis system (n = 4,428). Average SA, LA, FA, and FESR were 350 cm2, 220 cm2, 8.7 cm2, and 2.5%, respectively. Variability of FA (CV = 42%) and of FESR (CV = 39%) was four times greater than that of SA and LA. Slaughter group, pig, operator, and operator x pig effects were the most (P < 0.01) important sources of variation of measures. Correlations between measures obtained with different techniques were greater (P < 0.01) than 0.90, with the exception of LA measures. Coefficients of reproducibility for SA and LA ranged from 87 to 94%, whereas those for FA and FESR ranged from 88 to 98%. Coefficients of repeatability ranged from 92 to 99%. Automatic-assisted and manual methods provided more reproducible and repeatable measures than the automatic technique. Spatial calibration of the software system was a key issue affecting reproducibility and repeatability. Reinstructing the analysts for spatial calibration enhanced both reproducibility and repeatability of all methods of analysis. Computer image analysis is a technique suitable for measuring lean and fatty areas in cross-sectioned hams, providing reproducible and repeatable measures, and it might be used in large sample-based studies to investigate causes of defective fatty areas.

Adipose Tissue↗
PubMed2006

Fluorescent image analysis of lipid hydroperoxides in fish muscle with 3-perylene diphenylphosphine.

A fluorescent image analysis method was developed to evaluate lipid hydroperoxide formation in fish muscle. The lipid hydroperoxides generated in white and dark fish muscles during storage at 5-6 degrees C oxidized 3-perylene diphenylphosphine located in the tissue to yield the fluorescent derivative, 3-perylene diphenylphosphine oxide (3-PeDPPO). 3-PeDPPO thus obtained was determined by digital fluorescent image analysis. The 3-PeDPPO fluorescence intensity of white and dark muscle increased during low-temperature storage (0-24 h) and was clearly correlated with total lipid hydroperoxide levels in muscle extracts, which were determined by using HPLC based on a triphenylphosphine oxidation method (R2 = 0.954). These results suggest that 3-PeDPPO fluorescence, coupled with fluorescent image analysis, is a novel tool for direct determination of lipid hydroperoxides in fish muscle without a need for extraction of lipid.

Animals↗
PubMed2001

Prediction of principal metal-binding solid phases in estuarine sediments from color image analysis.

A novel application of image analysis technology to the semiquantitative determination of the strong metal-binding phases of hydrous ferric oxides and acid-volatile sulfides in muddy estuarine sediments is presented. Sediment cores were split in the laboratory, photographed in cross-section, and subsampled for solid-phase analyses. The images were processed using commercially available software packages. Concentrations of FeOOH and AVS were found to be linearly related to elementary color parameters (saturation and intensity, respectively) at corresponding points on the images. Spatially resolved information on metal-binding capacity can thereby be estimated from such images; selected depth profiles are generated as examples.

Color↗
PubMed1993

Quantitative analysis of gel electrophoretograms by image analysis and least squares modeling.

A computer-aided quantitative method for a complex analysis of gel electrophoretograms is presented. The analysis consists of several steps: (i) determination of the background image by methods of mathematical morphology and its subtraction from the gel image, (ii) selection of an appropriate part of the gel lane including curved lanes and lanes with a nonuniform width, (iii) computation of the lane densitogram by averaging several lane-parallel scans, (iv) decomposition of the lane densitogram into component bands using a data selecting algorithm and Marquardt's minimizer. Several different functions for component bands are utilized. It is shown that the densitogram can be decomposed into component bands with reasonable accuracy only if an appropriate model function is chosen. The algorithms are tested on several different gel electrophoretograms which show typical features as a nonuniform background, curved lanes, an asymmetrical band shape and a superposition of small bands on the shoulders of big ones. It is shown that overlapped bands are best approximated by an asymmetrical Gausian curve and an asymmetrical Gauss-Cauchy function. Linear response to the serial dilution of the protein sample is tested.

DNA, Bacterial↗
PubMed2006

[Examination of the increased speed of detection of HER-2/neu gene amplification in breast cancer by fluorescence in situ hybridization (FISH) using image analysis software].

The time required to count signals in the detection of HER-2/neu gene amplification in breast cancer by fluorescence in situ hybridization (FISH) has been a problem. To assess whether the amount of time necessary for counting could be reduced, image analysis using computer software (Win ROOF) was tested. Five photographs from each FISH sample were arranged into ten composite photographs. All ten composite photographs were necessary when using the conventional method of manual counting. However, using only four of the composite photographs and the image analysis method, the 60 necessary nucleus numbers could be measured, and a constant ratio of HER-2/neu / CEP 17 was obtained. In all 58 samples used, in the presence or absence of HER-2/neu gene amplification, there was agreement in counts between the conventional and image analysis methods, and a good correlation of r=0.961 (p<0.001) was obtained. Using the image analysis method, the necessary scoring time was reduced, particularly when the HER-2/neu gene had been amplified, where it was completed in about 1/4 of the time normally required. These results indicate that this image analysis method can be applied when using FISH in other areas of research, and may increase the speed of examination.

Breast Neoplasms↗
PubMed1993

Quantitative measurement of hemosiderin deposition in tissue sections of the liver by image analysis.

Hemosiderin has been quantitated in tissue sections by image analysis employing Perls' Berlin blue reaction. A linear correlation was observed between the mean optical density of the tissue section measured with a commercial image analyzer and the iron concentration measured with an atomic absorption spectrophotometer. The correlation coefficient of .945 (P < .001) indicated that a linear correlation existed between these measurements. Thus, the hemosiderin volume in tissue sections can be measured quantitatively by image analysis.

Hemosiderin↗
PubMed1986

An image analysis system for the determination of tooth dimensions from study casts: comparison with manual measurements of mesio-distal diameter.

A first comparison of an image analysis method with classical hand measurement was made using the mesio-distal diameter data derived by the simplest image analysis method, the jawview mode. Fifty casts of male Hong Kong Chinese dental students, mean age 20.4 years, were examined by both methods. The two techniques gave comparable overall results, the correlation coefficients generally being greater than 0.9. A combination of the buccal and occlusal image analysis views gave the best correlation with hand measurement with a comparable variability for different tooth types. For certain teeth, particularly posteriors, reproducibility was better using hand measurement. Optimal weighting of the data from these two different image analysis views can give an "estimated hand measurement" value. Further development of the method to deal with differing orientations of teeth is possible.

Adult↗
PubMed2002

Quantitative immunohistochemical evaluation of MIB-1 labeling index in adult soft-tissue sarcomas by computer-assisted image analysis.

We have found that the MIB-1 grade, based on tumor differentiation/histological type, necrosis and Ki-67 (MIB-1) score, is a valid and reproducible prognostic system for adult soft-tissue sarcomas. However, there are limited data available on Ki-67 labeling indices (LI) from adult soft-tissue sarcomas for testing the validity of quantitative image analysis. In this study, the records of 146 adult patients with soft-tissue sarcomas of the extremities and trunk were retrieved, and MIB-1 immunostaining was carried out for the grading. The counted MIB-1 LI values and the scores estimated from microscopic observation were defined as the gold standard. The correlation between MIB-1 LI as assessed by computer-assisted image analysis and by microscopic observation was determined. The image analysis -based MIB-1 LI was highly correlated with the microscopic observation-based MIB-1 LI (r = 0.87, 95% confidence interval (CI) = 0.82-0.92). In addition, agreement between the MIB-1 scores was very high (kappa statistic = 0.83, 95% CI = 0.75-0.91), as was the percentage agreement (89%, 95% CI = 82.8-93.6%) between the results from image analysis and microscopic observation. We conclude that quantitative immunohistochemical evaluation of MIB-1 LI by image analysis enables pathologists to improve interobserver agreement in the assessment of MIB-1 score, and can help to objectively assign the correct histological grade to cases of adult soft-tissue sarcoma, resulting in optimal clinical management.

Adult↗
PubMed1987

Measurement of radiographic changes occurring in rheumatoid arthritis by image analysis techniques.

We have applied image analysis techniques to serial measurements of bone contour in standard radiographs of single small joints of the hands in subjects with rheumatoid arthritis (RA) and controls. Adequate reproducibility was shown in 20 controls radiographed twice over a six month period. The technique showed significant changes in a proximal interphalangeal joint of 13 of 15 patients with RA studied over periods of three to 10 months. In further serial studies in selected small joints of RA hands significant changes could be shown as early as four months. These results justify further development of these techniques to allow their full scale evaluation in multiple joints in patients with RA receiving long term drug therapy.

Adult↗
PubMed1986

Measuring S100 protein and neurone specific enolase in melanocytic tumours using video image analysis.

Using a computed video image analysis system, the staining intensity for both neurone specific enolase (NSE) and S100 protein was measured in sections from 19 malignant melanomas and 16 benign melanocytic lesions. The results of this study confirm previous reports that NSE and S100 protein are useful markers for malignant melanoma. NSE staining intensity in the cases of malignant melanoma was significantly higher than that in benign naevi (p = 0.011). Intensity of staining for S100 protein was not significantly higher in the malignant melanomas. There was, however, a significant S100 gradient when comparing superficial and deep intradermal portions of these tumours (p = 0.003). This feature was not seen in benign naevi. The greatest intensity of S100 protein staining was found in the deeper portions of the malignant melanomas. This gradient difference was not seen with staining for NSE. Although it seems that the overall intensity of staining for NSE is more effective in differentiating between benign and malignant lesions, the difference in staining intensity between the superficial and deep portions of the tumour may be the better indicator of adverse behaviour in lesions in which the diagnosis of malignancy is uncertain.

Humans↗
PubMed1994

Ploidy pattern and cell cycle in breast cancer as detected by image analysis and flow cytometry.

Both image analysis (IA) and flow cytometry (FCM) may be applied to detect ploidy pattern and cell cycle fractions. However, they have different performance characteristics and may yield different results. The two approaches are applied in this study to 66 breast cancers: IA on imprints and FCM on fresh tissue. The percent coefficient of variation (CV) ranged from 2.0 to 7.0 (mean 5.5; SD 1.1) in IA and from 2.0 to 7.0 (mean 4.4; SD 1.1) in FCM. The values were well correlated. With regard to ploidy pattern, the agreement between the two methods was 92.4%; disagreements were due to four cases being aneuploid by IA but not detected by FCM and one case being aneuploid by IA but tetraploid by FCM. This suggests that IA is capable of detecting aneuploidy with more sensitivity than FCM. In diploid cases, the percent values of cells in G0/G1, S-phase (SPF), and G2M phase were concordant and well correlated. In aneuploid cases, IA was more sensitive than FCM in detecting aneuploid fraction as well as G2M phase, whereas FCM was more sensitive than IA in detecting SPF. A good correlation was found between the DNA indexes (DIs) obtained with the two methods.

Aneuploidy↗
PubMed1997

Tumor cell counting using an image analysis program for MIB-1 immunohistochemistry.

A cell counting method for MIB-1 immunohistochemistry, using an image analysis program, NIH Image, on a personal computer, has advantages over manual cell counting by microscopy. MIB-1 slides were photographed at a magnification of x 50 on 24 x 36 mm color films and the photographs were then enlarged to adequate size for observation. The MIB-1-positive cells, on the enlargements, were marked with a white pen, and negative cells with a black pen. Subsequently, the image of an enlargement was converted into data that can be processed by the program using a flatbed image scanner. Threshold levels were adjusted according to the respective white and black markings, and the program automatically counted the number of MIB-1-positive and negative tumor cells. Our method has the following significant advantages: Once cells had been marked and the image converted into the NIH Image data, the cell count process was completed within a few minutes; in contrast to manual microscopic cell counting, count overlapping or cells missed by the observer are eliminated; overlapping nuclei could be separated by the marking procedure; our method required no elaborate and costly image analysis system. The present method may be recommended to researchers undertaking cell kinetic studies utilizing immunohistochemical methods.

Brain Neoplasms↗