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Results for “Image Cytometry”

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At least 1,675 records · Page 93Linked to original sources
PubMed1999

Prognostic significance of DNA image cytophotometry for osteosarcoma.

OBJECTIVE: To investigate the prognostic significance of DNA image cytophotometric data. STUDY DESIGN: Twenty-six osteosarcomas in patients without lung metastases were investigated for several cytophotometric data. In 24 cases, these data were correlated with the clinical course of the patients to assess the prognostic value of nuclear DNA content in osteosarcomas. RESULTS: Of all osteosarcomas, 96% showed aneuploid DNA content. Patients with tumors having a 2c deviation index (2cDI) of 12.00, DNA malignancy grade (DNA-MG) of 2.0, a mean DNA content (MDC) of 4.95 c, DNA index (DI) of 1.75 or mean nuclear area (MNA) of 130 microns 2 had a significantly lower overall survival rate as compared to those with lower values (P < .05). CONCLUSION: Image cytophotometric features, such as 2cDI, DNA-MG, MDC, DI and MNA, are of prognostic value in patients with osteosarcoma and free of lung metastases.

Adolescent↗
PubMed1999

DNA content by in situ fluorescence imaging and S-phase detection, with chromatin structure preserved.

OBJECTIVE: To test the feasibility of in situ DNA quantitation of adherent cells' nuclei by fluorescence imaging, preserving chromatin structure and to follow-up S phase, in relation to DNA content, in order to assess the precision of DNA measurements. STUDY DESIGN: Double labeling experiments involved total DNA staining with Hoechst 33342 and BrdU immunostaining (after either Br photolysis and DNA strand break labeling by terminal transferase or acid denaturation) to detect replicating DNA. An epifluorescence microscope was used, images captured with a CCD camera and quantitative total DNA measurements done in 12 bits with IPLab software. BrdU results were related to DNA content on an individual cell basis. Cell cycle analyses were run with Imastat software (developed in the laboratory) on Hoechst-stained cells and on double labeled cells. RESULTS: In cells progressing through the cycle, as assessed by BrdU, a corresponding increase in DNA content was measured. Early S differed from G1 (P < .05). Imastat analyses gave a CV for GI peak of 6-7%. CONCLUSION: Quantitative fluorescence imaging allows a sensitive determination of DNA content for adherent-cell nuclei in situ. Topologic analyses of nuclear components will be possible in relation to DNA content.

3T3 Cells↗
PubMedDate not supplied

Image cytophotometric DNA analysis of cystadenolymphomas.

Warthin's tumour of the parotid gland is generally a benign lesion, consisting of an epithelial and a lymphoid component with unclear histogenetic relationship, 17 cases of Warthin's tumour were investigated for ploidy related parameters by the image analysis system CAS200. All cases investigated were euploid. A diploid DNA pattern (Auer type I) was detected in 7/17 (41%) cases both in the epithelial and lymphoid component of the same tumour. In addition to a diploid cell fraction a tetraploid cell component was found in 7/17 (41%) cases, coincidentally, in the epithelial and lymphoid component (Auer type II). Euploidy in all investigated cases is in agreement with the generally benign nature of Warthin's tumours. The similarity of the DNA pattern (Auer type) in the genetically unrelated epithelial and lymphoid components in most cases (84%) is suggested to be related to a similar functional state of these two components and is discussed in relation to the histogenesis of cystadenolymphomas.

Adenolymphoma↗
PubMedDate not supplied

Changes in DNA content of myocardial cells after cardiac explantation.

To evaluate the changes in DNA content of myocardial cells, image cytometric measurement was performed on a series of specimens obtained from 7 explanted hearts with respect to different fixation times after cardiac explantation. Prior to fixation, the tissue samples were stored at 4 degrees C or at room temperature. When the tissue samples were stored at room temperature, the integrated optical density decreased after 48 hours from cardiac explantation. Meanwhile, the coefficient of variation of integrated optical density and the sum of intermediate ploidies of myocyte nuclei increased. However, no significant changes were found when tissue samples were kept at 4 degrees C. This study indicates that no significant changes in DNA content are found within two days from cardiac explantation.

Cardiomyopathies↗
PubMed1999

Appearance of highly sensitive cells to L-type Ca antagonist in cultured cerebellar granule cells.

We studied intracellular Ca2+ ([Ca2+]i) kinetics within single cells using and image-analyzing system. We prepared a culture dish with a bottom made of quartz (non-fluorescent glass) for reducing the background fluorescence during [Ca2+]i measurement. Furthermore, we changed the concentration of the poly-L-lysine solution which had been used to make cells adhere to the dish bottom from the conventional 5 micrograms/ml to 15 micrograms/ml. This modification improved cell viability, prevented colony formation, and therefore, provided favorable cell samples for detection within single cells. Then, we investigated 25 mM KCl-evoked changes in the [Ca2+]i level of rat cerebellar granule cells. We found that sensitivity to KCl was high in some cells but low in others. In a study using the highly sensitive cells, nicardipine, nimodipine and nifedipine were active even at nanomolar or picomolar levels, although their affinities to the L-type Ca ion channel have been reported to act only at micromolar levels.

Animals↗
PubMed2000

[Morphometric and cytogenetic characteristics of haploid tomato plants].

Stable differences have been found between haploid and diploid Mikado tomato plants with respect to quantitative characteristics of their vegetative and generative organs, whereas these plants were phenotypically similar with respect to qualitative characteristics, such as the bush habitus and the shapes of leaves, flowers, and fruit. A simple indirect method based on counting the chloroplasts in guard cells has been suggested to diagnose haploids. Analysis of meiosis during microspore formation in haploids has revealed considerable anomalies leading to heterogeneity and a high sterility rate of pollen, which has been confirmed by differences in DNA content. The data obtained suggest that formation of fertile gametes is accounted for by the absence of reduction division. It has been hypothesized that meiosis in haploids induces the genotypic variation that is subject to selection.

Cytogenetics↗
PubMed2000

Affinity of linker histones for chromatin in situ analyzed using DAPI as a cytochemical probe.

BACKGROUND: It is generally assumed that linker histones contribute to condensation of chromatin and to the repression of genetic activity by binding to chromatin with increased affinity. The aim of this study is to investigate a possible correlation between linker histone binding to chromatin in situ and chromatin condensation using a cytochemical approach that largely preserves nuclear structure. METHODS: Ionically bound H1 was extracted with increasing concentrations of sodium chloride. Thereafter, the cells were fixed in formaldehyde and stained with 50 nM 4',6-diamidino-2-phenylindole (DAPI) and fluorescence intensity was measured by image cytofluorometry. RESULTS: The association between linker histones and chromatin was stronger in cultured human fibroblasts and rat smooth muscle cells than in both human T lymphocytes and granulocytes, and in particular, frog erythrocytes, which exhibited highly condensed chromatin. Our data indicate a lower affinity of linker histones for metaphase chromosomes than for interphase chromatin, and we observed a reduction in linker histone affinity during terminal differentiation of frog erythrocytes in vivo. CONCLUSIONS: Taken together, our findings imply that the affinity of linker histones for chromatin in situ is unrelated or inversely related to chromatin condensation.

Animals↗
PubMed2000

Quantitative cytology in ovarian carcinoma ascitic fluids.

OBJECTIVE: To assess the value of quantitative methods in the differential diagnosis between ovarian carcinoma cells and mesothelial cells in ascitic fluids. STUDY DESIGN: Ninety ascitic fluid samples, previously reported as positive for ovarian carcinoma (30 cases), suspicious for malignancy (30) and negative for malignancy, containing only reactive mesothelial cells (30), were retrieved from the files. In each of these specimens the nuclear area, perimeter, roundness and shape coefficient of 100 cells were determined at 630 x magnification. Statistical analysis was performed using analysis of variance and, for multiple comparisons, the Student-Newman-Keuls technique. RESULTS: Mean values for nuclear area and perimeter were higher in malignant cells as compared to reactive mesothelial cells, whereas those for roundness and shape coefficients were lower. All differences were statistically significant, the former two at a .05 level and the latter at the .001 level. CONCLUSION: Quantitative methods can reliably support the differential diagnosis between ovarian carcinoma cells and mesothelial cells in ascitic fluid specimens.

Aged↗
PubMed2000

[Cytological and cytometric characteristics of morphologic variants of thyroid papillary cancer].

A total of 570 cases with thyroid papillary cancer are analyzed. Eight cytologic variants of the tumor were distinguished: typical (classical), 240 (42.1%) cases; follicular, 225 (39.5%); cystic, 70 (12.3%); with squamous-cell metaplasia, 16 (2.8%); with B-cell transformation, 10 (1.7%); from high cells, 6 (1.0%); anaplastic, 2 (0.4%); and sclerosing, 1 (0.2%) cases. Cytometric analysis of tumor smears from 103 patients was carried out. Four parameters were acknowledged as the most valuable for verification of thyroid papillary cancer variants: cell perimeter, cell area, nuclear area, and nucleus/cytoplasm ratio. Cytologic and cytometric findings allow the clinical cytologist verify the variant of thyroid papillary cancer in the majority of cases.

Adult↗
PubMed2000

Using fos imaging in the rat to reveal the anatomical extent of the disruptive effects of fornix lesions.

Activity of the immediate early gene c-fos was compared across hemispheres in rats with unilateral fornix lesions. To engage Fos production, rats first performed a radial arm maze task that is severely disrupted by bilateral fornix lesions. Using immunohistochemical techniques, Fos-positive cells were visualized and counted in 39 sites in both hemispheres. Fornix lesions led to a significant reduction in Fos in all ipsilateral hippocampal subfields, as well as the entorhinal cortex and most of the subicular complex. Other sites that showed reduced activity included the ipsilateral retrosplenial, anterior cingulate, and postrhinal cortices. Subcortical regions showing significant Fos decreases included the anterior thalamic nuclei, supramammillary nucleus, diagonal band of Broca, and lateral septum. Thus, the effects of fornix lesions extended beyond the hippocampal formation and included sites not directly innervated by the tract. These changes were nevertheless selective, as shown by the lack of hemispheric difference in any of the preselected control sites, the perirhinal cortex, or nucleus accumbens. Furthermore, there were no hemispheric differences in an additional group of animals with unilateral fornix lesions that were killed directly from the home cage. The location of Fos changes closely corresponded to those brain regions that when lesioned disrupt spatial working memory. Moreover, there was a correspondence between those brain regions that show increased Fos production in normal animals performing the radial arm maze task and those affected by fornix lesions. These results show that fornix transection has widespread, but selective, effects on a network of structures normally activated by spatial memory processes, with these effects extending beyond the hippocampal formation.

Animals↗
PubMed2001

Expression of the Kv3.1 potassium channel in the avian auditory brainstem.

The Shaw-like potassium channel Kv3.1, a delayed rectifier with a high threshold of activation, is expressed in the time coding nuclei of the bird auditory brainstem. In both barn owls and chickens, Kv3.1 mRNA was expressed in the cochlear nucleus magnocellularis (NM) and the nucleus laminaris (NL). Western blot analysis showed that an antibody raised against the synthetic peptide sequence of rat Kv3.1 (rKv3.1) specifically recognized the same 92 kDa protein bands in both rat and chicken synaptosomal preparations. Immunohistochemical analyses using this anti-rKv3.1 antibody revealed a prominent gradient in Kv3.1 immunoreactivity along the tonotopic axis of the barn owl NM and NL and a less prominent gradient in the chicken. The precise localization of the Kv3.1 immunoproduct was resolved by electron microscopy. In both the owl and the chicken, Kv3.1 was targeted postsynaptically in NM and NL. The major difference in localization of Kv3.1 protein between the two birds was the expression of Kv3.1 in the NM axons and terminals in the region of the barn owl NL. This location of Kv3.1 channels supports its postulated function in reducing the width of action potentials as they invade the presynaptic terminal. The presynaptic localization may be a specialization for enabling neurons in owl NM to transmit high-frequency temporal information with little jitter.

Alternative Splicing↗
PubMed2001

Progression curves for endometrial lesions.

OBJECTIVE: To derive a numeric measure for the progression of endometrial lesions as a baseline study for an eventual assessment of chemopreventive intervention efficacy. STUDY DESIGN: Tissue sections from normal endometrium at the proliferative and secretory phase, simple hyperplasia, atypical hyperplasia from cases free of concomitant adenocarcinoma and adenocarcinoma of the endometrium were recorded at high spatial resolution. Six cases from each diagnostic category were chosen as "typical," and 60 epithelial nuclei were randomly selected for measurement for each case. Discriminant analyses were carried out to derive a direction of progressive change in feature space and to correct the progression curve for the presence of cells not expressing progressive change among the random sample of nuclei. RESULTS: A well-conditioned progression curve was derived based on the mean discriminant function scores for each diagnostic category and the mean nuclear abnormality of the nuclei in each category, as expressed by their deviation in feature values from normal reference nuclei. The lesion signatures showed a clear trend toward extension into the range of higher nuclear abnormalities with increasing progression. There was an indication that abnormal endometrial lesions may comprise cases with distinctly different degrees of nuclear abnormality. CONCLUSION: A numeric assessment of lesion progression for endometrial lesions, based on karyometric measurements, is possible. The data suggest that additional analysis may provide further characterizing information for individual lesions.

Adenocarcinoma↗
PubMed2001

Microvessel density counting in breast cancer. Slides vs. digital images.

OBJECTIVE: To develop a program to assist the pathologist in the acquisition and evaluation of digital images to determine microvessel density (MVD) in tissues. STUDY DESIGN: Ten cases of breast cancer with a high degree of neovascularization were selected. A standard immunohistochemical method was used to highlight the microvessels (monoclonal anti-factor VIII, avidinbiotin-peroxidase complex method). Two pathologists (one senior [S] and one junior [J]) evaluated four areas of highest neovascularization ("hot spots") in the tumors. Microscopically MVD was determined in four chosen areas (400:1). From the center of each area two digital images were acquired at a magnification of 200:1. All counts made by microscopic observation were compared with those made on the digital images. To compare MVD counting at different resolution, two sets of images at different sampling densities (320 x 240 and 1,600 x 1,200) were assessed by the two pathologists. RESULTS: We obtained a good correlation (r = .98 for S and .96 for J) between the MVD counts obtained at the microscope (192.8 MV/mm2 [mean of S] and 181.8 MV/mm2 [mean of J]) and the MVD counts from digital images (153.2 MV/mm2 [mean of S] and 171.0 MV/mm2 [mean of J]) at high resolution. The counts were lower for digital images at lower sampling density (125.0 MV/mm2 [mean of S] and 78.2 MV/mm2 [mean of J]). With low-resolution digital images only S maintained a good correlation (r = .96 for S and .34 for J) with the microscopic evaluation of MVD. Interobserver analysis showed a good correlation (r = .82 for the microscope and r = .78 for the digital images) of MVD evaluated either at the microscope or in high-resolution digital images. CONCLUSION: We demonstrated the functionality and usefulness of our program in performing MVD evaluation. Considering the capabilities of the program to store all images and microvessel marks and the reliability of MVD evaluation based on digital images, we consider this program the first step toward fully automated MVD assessment.

Breast Neoplasms↗
PubMed2001

Karyometry in Barrett's esophagus.

OBJECTIVE: To derive a progression curve for lesions in Barrett's esophagus based on karyometric features. STUDY DESIGN: High-resolution imagery of 900 nuclei from normal gastric tissue, Barrett's metaplasia, Barrett's high grade dysplasia and adenocarcinoma of the esophagus was recorded. Karyometric features were computed, and nuclear signatures and lesion signatures for these lesions were derived. A progression curve was defined. RESULTS: Esophageal lesions were distinctly different from the normal gastric fundus tissue, with nuclei from Barrett's metaplasia deviating from normal almost as much as nuclei from high grade dysplasia and adenocarcinoma. There was considerable case-to-case variability and overlap between lesions histologically assigned to different diagnostic categories. CONCLUSION: The karyometric data suggest that Barrett's metaplasia is a more developed lesion than previously assumed.

Adenocarcinoma↗
PubMedDate not supplied

[Morphometric and immunohistochemical markers of breast cancer progression].

The results of the study for the last decade aimed at the search of markers of mammary carcinoma malignancy and prognosis are reported. Accumulation and integration of various data contributes not only to more precise identification of the malignancy but also to more complete understanding of biological essence of mammary carcinoma.

Biomarkers, Tumor↗