Evaluation of LD BC-Ⅰ Blood Cell Image Automatic Analyzer in Analysis of the Nucleated Cells in Peripheral Blood Smear / 现代检验医学杂志

Objective To verify and evaluate the clinical performance of LD BC-Ⅰ blood cell image automatic analyzer.Methods A total of 202 EDTA-Na2 anticoagulant blood specimens were collected at the hospital clinic randomly between October and December 2016.After wright staining,each specimen was examined by microscopy and automatic analyzer respectively,the detection efficiency and the results of nuclear cells classification were compared between artificial microscopy and automatic analyzer,the correlation and consistency of two methods were further analyzed.Results The average time required for each specimen of the automatic analyzer was reduced by 3.81 minutes when compared with artificial microscopy,the P values was less than 0.01.LD BC-Ⅰ agreed 71.4％,64.8％,28.8％,21.1％ and 71.6％ respectively for pre-differentiation of the blast cells,promyelocytes,myelocyte,metamyelocytes and atypical lymphocytes.The diagnosis accordance rates of five nucleated cells above increased to 87.7％,81.5％,38.1％,26.3％ and 86.2％ after manual review.Passing Bablok regression analysis found that the correlation coefficient (r) of two methods of neutrophils was 0.981,lympho kjcytes (r =0.894),monocytes (r=0.725),eosinophils (r=0.772),and there were significant correlation between two methods (all P ＜0.01).Bland-Altman analysis found that the coincidence rate of the neutrophils and lymphocytes were 96％,the monocytes were 91％ and the eosinophils were 94％.Conclusion The LD BC-Ⅰ automatic blood cell image analyzer could significantly improve the analysis efficiency of the nucleated cells.There was good consistency in the classification results of mature granulocytes,monocytes and lymphocytes between instrumental detection and artificial microscopy.The method has certain clinical value to be applied widely.

Performance of an Automated Fluorescence Antinuclear Antibody Image Analyzer

BACKGROUND: The gold standard for antinuclear antibody (ANA) screening is the indirect immunofluorescence (IIF) assay with human epithelial cells (HEp-2). However, a number of substantial disadvantages of manual IIF assays have highlighted the need for the automation and standardization of fluorescent ANA (FANA) testing. We evaluated the performance of EUROPattern Suite (Euroimmun AG, Germany), an automated FANA image analyzer, with regard to ANA detection and pattern recognition compared with conventional manual interpretation using the fluorescence microscopic IIF assay. METHODS: A total of 104 samples including 70 ANA-positive sera and 34 ANA-negative sera collected from September to October 2015 were included. The sensitivity, specificity, and pattern recognition function were evaluated to determine the performance of EUROPattern Suite compared with the manual IIF assay results. RESULTS: The sensitivity and specificity of EUROPattern Suite for ANA detection were 94.3% and 94.1%, respectively. The concordance rate between the two methods was 94.2%. For pattern recognition, 45.7% of the samples were assigned identical ANA patterns including simple and mixed. When major pattern matching was considered, 83.7% (41/49) and 95.2% (20/21) of the samples with simple and mixed patterns, respectively, showed concordant results between the two methods. CONCLUSIONS: EUROPattern Suite, an automated FANA image analyzer, provides a viable option for distinguishing between positive and negative results, although the ability to assign specific patterns is insufficient to replace manual microscopic interpretation. This automated system may increase efficiency in laboratories, in which a large number of samples need to be processed.

A comparison of marginal fit of glass infiltrated alumina copings fabricated using two different techniques and the effect of firing cycles over them

PURPOSE: This study evaluated marginal fit of glass infiltrated alumina cores fabricated using two techniques and their marginal stability after firing cycles of veneering porcelain. MATERIALS AND METHODS: Fifteen standardized all-ceramic crowns were fabricated on a metal die using each technique: slip cast technique of VITA In-Ceram sprint Alumina (Group A as control) and plastic foil matrix technique of Turkom-Cera fused alumina core system (Group B). Copings were compared between groups and within groups at coping stage and after firing each layer of veneering porcelain. A device was used to standardize seating of copings on the metal die and positioning of the specimens under the microscope after each stage of fabrication. The specimens were not cemented and marginal gap was measured using an image analyzing software (Imagepro Express) on the photographs captured under an optical microscope. Two tailed unpaired 't test' was used to compare marginal gaps in two groups and one way ANOVA was used to analyze marginal distortion within each group at 95% confidence interval. RESULTS: The marginal gap was smaller at the coping stage in group B (60 + 30 microm) than group A (81 + 21 microm) with statistical significance. After firing of veneering porcelain the difference was insignificant. At the final stage, both groups exhibited lower mean marginal gaps than at the initial coping stage with the difference of 11.75 microm for group A and 11.94 microm for group B, but it was statistically insignificant due to high value of standard deviation. CONCLUSION: Within the limitations of this study, it was concluded that both techniques produced copings with comparable and acceptable marginal fit and marginal stability on firing veneering porcelain.

A Comparison of Acquired Port-wine Stain with Congenital Port-wine Stain Using an Image Analyzer

BACKGROUND: Recent reports have proposed that there were no differences between acquired port-wine stain (APWS) and congenital port-wine stain (CPWS) except the onset of disease. Pulsed dye laser (PDL) therapy is regarded as the treatment of choice in PWS. Although in some articles, APWS might have shown a better response to PDL than CPWS, this is still controversial. It has been assumed however, that there might be some differences determining therapeutic responses between the two entities. OBJECTIVE: The purpose of this study is to find out some histopathologic differences between APWS and CPWS. METHODS: 14 patients with APWS and 17 patients with CPWS from our patient files were included in this study. Immunohistochemical staining by factor VIII-related antigen was carried out on the specimens of punch biopsy to better visualize the blood vessels. Histopathologic assessment of variables such as vessel area, percentage of vascular area and vessel depth was performed using a computer-assisted image analyzer program. RESULTS: The mean vessel area in APWS was 1014.7 +/- 782.5micrometer2 and that of CPWS was 1341.5 +/- 689.9micrometer2. The mean percentage of vascular area in APWS was 2.02 +/- 1.38% and that of CPWS was 2.65 +/- 1.56%. The mean vessel depth in APWS was 327.5 +/- 120.7micrometer and 321.7 +/- 93.1micrometer in CPWS. No histopathologic variable was statistically significant using the Mann- Whitney test (p>0.05).

Therapeutic Effect of Multiple Paper Patching for Traumatic Tympanic Membrane Perforation-Trial of Quantitative Analysis Using Image Analyzer / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Paper patching for the treatment of traumatic tympanic membrane perforation is a safe, simple, and inexpensive method. This study was designed to evaluate the therapeutic effect of proper paper-patch techniques for traumatic tympanic membrane perforation using more objective measurement of perforated area by image analyzer. SUBJECTS AND METHOD: A prospective study of paper-patch techniques was carried out on 55 patients with traumatic perforations of the tympanic membrane. Objective semiquantitative measurement of the perforated area was performed using computerized image analyzer. Immediate eversion of inverted flaps and single to multiple patching techniques were used in all cases. Treatment results were analyzed according to various variables which may affect the healing rate. RESULTS: The closure rate of tympanic membrane with this paper patching method was 98.2% and the mean healing time was 2.9+/-1.9 weeks. Hearing impairment was correlated with the size of perforation which was measured quantitatively by image analyzer and the healing rate was decreased with infectious signs such as otorrhea. There were no side effects or complications. CONCLUSION: Multiple paper patching with proper technique for traumatic tympanic membrane perforation produced an excellent success rate and could be considered as part of initial therapeutic methods for all cases of large perforation with inverted flap.

Morphometric Analysis for Cytological Diagnosis of Thyroid Papillary Carcinoma / 대한세포병리학회지

The diagnosis of papillary thyroid cancer is generally based on the findings of intranuclear cytoplasmic inclusions and nuclear grooves. Although anisokaryosis and poikilokaryosis, in papillary thyroid cancer, are not distinct when compared to other cancers, cytological examination can provide useful preoperative information. Our study evaluated the diagnostic role of computer-assisted image analysis for the pre-surgical assessment of papillary thyroid carcinoma. Thyroid aspirates from twenty female patients who were histologically confirmed to have both papillary carcinoma and benign nodules were studied. Different populations of 50 benign cells and 50 malignant cells were analyzed. Five morphometric parameters were selected for analysis: nuclear area, perimeter, maximum length, maximum width and intensity standard variation. The values obtained for papillary carcinomas were higher than the surrounding benign nodules as follows: nuclear area 63.5 vs. 36.1 (p=0.000), nuclear perimeter were 29.4 vs. 22.0 (p=0.000), maximum length 9.6 vs. 7.1 (p=0.000), maximum width 8.2 vs. 6.3 (p=0.000), the ratio between maximal length and maximal width 1.16 vs. 1.13 (p=0.000), the standard variation of intensity 14.9 vs. 15.9 (p=0.101) respectively. Therefore, morphometric information can be helpful for the differential cytological diagnosis of papillary thyroid carcinoma.

Effects of Platelet-derived Growth Factor and Laminin on Retinal Pigment Epithelial Cells

To investigate the effects of platelet-derived growth factor(PDGF)and laminin on the proliferation and morphological change of retinal pigment epithelial(RPE)cells, RPE cells(5 10(3))were cultured on either plastic or laminin-coated 24-well-plates for 6 days. One percent fetal calf serum and PDGF(0, 10, 20 ng/ml)were added in the culture media. Morphological change and the numbers of attached RPE cells were assessed at the end of the culture, using image analyzer and MTT assay. In image analysis, as the PDGF concentration were increased, area(A), perimeter(P), normalized perimeter(NP), diameter(D), minimum size(MinS), maximum size(MaxS), and eccentricity of binary object(EF)were increased while circle filling(Cfil)was decreased. Compared with the cells cultured on plastic, the cells on the laminin showed increase in all the parameters except EF. The proliferative effect on RPE cells were increased on both plastic and laminin as PDGF concentration were increased(P<0.05). These results suggest that proliferation and morphologic change of RPE cells might be influenced by PDGF and laminin.

Apoptosis in Ventral Lobe of Rat Prostate after Castration: Morhometric Measurement to Epithelial Cell Area / 대한남성과학회지

PURPOSE: While apoptosis of the rat prostate cells has been the focus of many studies on androgen action and prostatic carcinogenesis, little is known for morphometric measurement about rat prostatic glands. Apoptosis was described as a morphologically distinct form of cell death. However, the approach of morphological assessment in one cell to define apoptosis has some disadvantages. We studied androgen ablation induced reduction of glandular epithelial cells with counting apoptotic cells and measuring glandular cell mass with image analyzer. MATERIALS AND METHODS: Male Sprague-Dawley rats were used. The morphologic changes were evaluated at 4, 7, 14 days after castration. The ventral lobe was microdissected. The distal part of it was harvested under guide of dissecting microscope. The number of apoptotic cells were quantified in Masson & trichrome stained specimens. Using image analyzer, area densities in glandular lesion of prostatic epithelium were calculated. RESULTS: After castration. the distal duct may seem to be melted down into the proximal ducts and tall columnar epithelial cells of distal duct seem to be changed into cubodial cells. The apoptotic counts were 1.00+/-0.28, 5.06+/-0.98, 6.01+/-0.97, and 2.04+/-0.70 in control, 4th, 7th and 14th day group, respectively (p<0.05), The areas of epithelial cell mass were 13874+/-3106 micrometer2. 5530+/-3244 micrometer2, 2345+/-1226micrometer2, and 1339+/-652 micrometer2. in control, 4th. 7th and 14th day group, respectively(p<0.05). CONCLUSIONS: We conclude that castration induces a more profound involution of the rat ventral prostate. After castration, the apoptotic count and reduction of the area of epithelial cell mass rapidly processed by 7th day and after that time, this process is continuously but slowly decreased until 14th day after castration. The apoptotic counts among the epithelial cells are well matched with the morphometric measurements about epithelial glands.

Morphometric Study of the Irradiation Effecton the Cartilage Formation in the Rat Mandibular Condyle / 대한구강악안면방사선학회지

PURPOSE: This study was undertaken to quantitatively estimate the degree of the damage and recovery of the irradiated rat condylar cartilage using the Image Analyzer. MATERIALS AND METHODS: Experimental animals were 16 male rats of the Sprague-Dawley strain at the age of 20 day irradiated with the dose of 10 Gy in their head and neck region. Four rats were sacrificed at the each of the

The Significance of DNA Image Analysis in Laryngeal Keratosis / 대한이비인후과학회지

Early diagnosis of laryngeal keratosis, a precancerous lesion, is important since it has the possibility to make a malignant change. Since it is believed that laryngeal keratosis progresses to carcinoma in situ or invasive carcinoma through the stage of epithelial proliferation and epithelial dysplasia, if we can predict this progression it will aid in the early diagnosis and treatment of laryngeal cancer. The authors used image analyzer to obtain the DNA index and proliferation index after DNA staining of keratotic epithelial cells to determine the probability of malignant change in laryngeal keratosis. We used specimen obtained from 38 cases of laryngeal keratosis. DNA ploidy was determined by obtaining DNA index with image analyzer(CAS 200, Image analyzer, CAS Ins, USA). And, proliferation index was obtained by ratio of cell content in S phase to G2/M phase of cell cycle. DNA ploidy in mild and moderate dysplasia showed diploidy in all cases, however, in four of nine cases of severe dysplasia showed aneuploidy. Cellular proliferation index in mild dysplasia was 7.5+/-3.12, in moderate dysplasia was 12.3+/-5.71, and in severe dysplasia was 15.8+/-4.21, and statistically significant(p<0.05) high cellular proliferation index was observed in relation to pathological classification. Therefore, DNA analysis using image analyzer can be used to determine the degree of progression of laryngeal keratosis, and it is thought as an objective method for predicting the probability of malignant change and prognosis of the patient.

Antiangiogenic Effects of Retinoids on Corneal Neovascularization

We investigated the effect of topical application of retimids on the corneal neovascularization in the rat induced by chemical cauterization. The center of corneas of Wistar rats were Cauterized with a silver/potassium nitrate applicator for 5 seconds. And then they were treated topically with 0.1%, 0.2%, 0.5%, 2.0% all-trans retimids (retinol, retinoic acid, retinaldehyde) and dimethyl sulfoxide as a control four times a day. After appiications of eye drops for 5 days, each rat was kined and then perfused with a mixture of 11% gelatin 10% India ink-lactated Ringer's solution. Corneal flat preparation were made, and then the percent of the corneal area occupied by blood vessels were analized by computerized image analyzer. Percent vascularization of 0.1% and 0.2% retnoids were not significantly different from control group(p>0.05). Percent vascu1arization of 0.5% retinaldehyde and all 2.0% retnoids were significantly lower than the contml group(p>0.5). These naturally occurring retinoids were effective in antiarlgiogenesis of rat cornea at relatively high concentrations (>0.5%), when treated topicany, It will be necessary to further study on more potent antiangiogenic, at lower concentration, synthetic retimids for the treatment of angiogenic diseases.

Antiangiogenic Effects of Retinoids on Corneal Neovascularization

We investigated the effect of topical application of retimids on the corneal neovascularization in the rat induced by chemical cauterization. The center of corneas of Wistar rats were Cauterized with a silver/potassium nitrate applicator for 5 seconds. And then they were treated topically with 0.1%, 0.2%, 0.5%, 2.0% all-trans retimids (retinol, retinoic acid, retinaldehyde) and dimethyl sulfoxide as a control four times a day. After appiications of eye drops for 5 days, each rat was kined and then perfused with a mixture of 11% gelatin 10% India ink-lactated Ringer's solution. Corneal flat preparation were made, and then the percent of the corneal area occupied by blood vessels were analized by computerized image analyzer. Percent vascularization of 0.1% and 0.2% retnoids were not significantly different from control group(p>0.05). Percent vascu1arization of 0.5% retinaldehyde and all 2.0% retnoids were significantly lower than the contml group(p>0.5). These naturally occurring retinoids were effective in antiarlgiogenesis of rat cornea at relatively high concentrations (>0.5%), when treated topicany, It will be necessary to further study on more potent antiangiogenic, at lower concentration, synthetic retimids for the treatment of angiogenic diseases.

Assessment of DNA Ploidy, Estrogen and Progesterone Recetor Status and Her-2/neu Oneoprotein Expression in Breast Carcinoma by Image Analysis

In 41 cases of breast cancers, the aneuploidy measured by Image Analyzer was compared with that of flow cytometric analysis, and estrogen and progesterone receptor(ER/PR) and Her-2/neu oncoprotein were immunohistochemically stained and measured by Image Analyzer. In ER/PR, the positive nuclear area(PNA, %) was measured, and in Her-2/neu, the content of oncoprotein was expressed as pg/cell. To assess the usefulness of these parameters as a prognostic factor, the author evaluated the results in relation with tumor size, nuclear grade and lymph node metastasis. The obtained results are summarized as follows: 1) The detection rate (90%) of aneuploidy by image analysis was higher than that (70%) of flow cytometric analysis. The concordance rate of both method was 80%. 2) The positivity of ER was 73% and PR was 34%, and the high PNA of ER and PR was related with high nuclear grade. There was an inverse correlation of the ER PNA with tumor size and PR PNA with negative lymph node. 3) Her-2/neu oncoprotein overexpression was found in only 2 cases and another two showed borderline overexpression. All four cases had DNA tetraploidy. From the above results, it was concluded that the image analyzer could be used in DNA analysis and in quantitation of immunostained ER/PR and Her-2/neu oncoprotein, providing the important information in the management of the breast cancer patients.