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At least 1,207 records · Page 67Linked to original sources
PubMed2000

Image analysis and flow cytometric DNA studies of benign and malignant body cavity fluids: reappraisal of the role of current methods in the differential diagnosis of reactive versus malignant conditions.

Cytologic examination of body fluids is commonly performed in the clinical laboratory. Determination of the presence of malignancy may sometimes be difficult. In this study, we prospectively studied 60 body fluids with a panel of antibodies, including MOC-31, epithelial membrane antigen, carcinoembryonic antigen, B72.3, keratin, desmin, and CA-125. DNA and S-phase studies were performed both by flow cytometry and image analysis. Thirty-seven fluids were classified as benign and 23 were classified as malignant. The sensitivity of the antibodies for identification of carcinoma in descending order of percentage detection rate were MOC-31 (95%), epithelial membrane antigen (93%), B72.3 (84%), and carcinoembryonic antigen (80%). Desmin stained mesothelial cells in all cases. CA-125 gave similar results but was less specific. Flow cytometry detected 14 of 20 malignant fluids and image analysis 17 of 23 by identifying an aneuploid population. Benign reactive mesothelial cells were not aneuploid. Tetraploidy due to reactive mesothelial cells was found in 9 of 37 body fluids. Their S-phase fraction was low (average, 3.2%). Tetraploidy in malignant cells was distinguished from the reactive mesothelial cells by high S-phase (average, 25.95). S-phase had some use as a discriminating factor, because no benign reactive cases had more than 17%. However, 7 of 23 malignant cases had a value below 17%. DNA analysis by image was more sensitive and specific than flow. Either may be used when immunocytochemistry is nondiagnostic or cannot be performed.

Adult↗
PubMed1992

Image cytometric DNA analysis in human breast cancer analysis may add prognostic information in diploid cases with low S-phase fraction by flow cytometry.

Measurements of DNA ploidy can be performed either with image cytometry (ICM) or flow cytometry (FCM); both methods provide independent prognostic information in primary breast cancer. The aim of the present investigation was to compare the two methods and to relate the findings to prognosis (median follow-up 42 months). Concordance in ploidy status (diploid, tetraploid, aneuploid) was obtained in 76% of the samples (168/222). When the fraction of S-phase cells (SPF) from FCM analysis was also taken into consideration, four different groups of samples were obtained (Flow I-IV), which were considered to correspond to the Auer classification (Auer I-IV) of DNA histograms obtained from image cytometry. Complete concordance between the two techniques now was 70% (155/222). Samples classified as Flow I (diploid or near-diploid with low SPF) and Auer I had a distant metastasis rate of 3/60 (5%), as compared to 62/154 (40%) for all other combinations of the Flow and Auer classifications taken together. Thus, the only findings of prognostic importance were that some samples were Flow I but not Auer I, or vice versa. These two groups represent 17 (7.7%) and 14 (6.3%), respectively, of the total number of samples, and had frequencies of distant metastasis similar to those of the other high-risk groups, namely, 7/17 and 5/14, respectively. In a multivariate analysis, flow cytometric S-phase value was a stronger prognostic factor than either the Flow and Auer classification. We conclude that when routine FCM DNA analysis is used, diploid or near-diploid samples with a low S-phase value should be reanalyzed with ICM.

Breast Neoplasms↗
PubMed1996

DNA ploidy and cell cycle analysis in cancer diagnosis and prognosis.

This review focuses on the clinical utility and potential value of cell cycle analysis and DNA ploidy interpretation in the diagnosis of human tumors, the application of these techniques to cytologic diagnosis, and their capability for predicting disease outcome in human neoplasia. Methods of cell cycle analysis are considered, and the techniques of flow cytometry and image analysis are described.

Cell Cycle↗
PubMed1991

Comparison of image analysis and flow cytometric determination of cellular DNA content.

A good correlation (r = 0.94) was obtained between the DNA indices (DI) using flow cytometry and image analysis of nuclei cytospins extracted from paraffin wax embedded tumour sections. Some of the limitations and problems associated with image analysis which came to light included an unacceptably high coefficient of variation (CV) and a "left-shift" in the DI in most DNA histograms obtained when using image analysis of 5 microns sections. In contrast, the DNA histograms generated using image analysis of cytospun nuclei from paraffin wax blocks were of good quality and similar to those obtained using flow cytometry. Variability in Feulgen staining was common and an important source of error despite rigorous control of the staining technique. This could be overcome by using internal controls such as fibroblasts rather than external controls (rat hepatocytes) to determine the diploid DI with image analysis. A thorough understanding and appreciation of the methodological problems associated with image analysis and flow cytometric determination of DNA content is required before these methods find widespread clinical application.

Adrenal Gland Neoplasms↗
PubMed1997

Primary malignant melanoma of the skin. Relationships of nuclear DNA content, nuclear morphometric variables, Clark level and tumor thickness.

OBJECTIVE: To investigate how nuclear morphometric variables, tumor thickness (measured according to Breslow), invasion depth (classified according to Clark), nuclear DNA content and type of DNA histogram are associated with each other in primary malignant melanomas of the skin. STUDY DESIGN: Image analysis DNA cytometry and nuclear morphometry were performed on 85 primary skin melanomas. The relationships of size, sphericity and DNA content of melanoma cell nuclei; melanoma thickness; and Clark level were analyzed in detail. The effect of melanin bleaching on DNA cytometry results was studied. RESULTS: Melanoma thickness correlated with nuclear size in aneuploid, but not diploid, melanomas. The prevalence of aneuploidy did not increase with tumor thickness. In aneuploid melanomas the proportion of cells with higher-than-diploid and higher-than-tetraploid DNA content increased with tumor size. CONCLUSION: Aneuploidy is as common in thin as in thick melanomas. Genetic instability in aneuploid melanomas correlates with melanoma thickness. This correlation in aneuploid melanomas partially explains the correlation between nuclear size and melanoma thickness. In diploid melanomas no correlation was observed between nuclear size and melanoma thickness. DNA cytometry is a valuable tool for studies on the background of phenotypic changes in skin melanomas.

Adult↗
PubMed2002

Exposure of cells to static magnetic field accelerates loss of integrity of plasma membrane during apoptosis.

BACKGROUND: Much attention is being paid to the biologic effects of magnetic fields (MFs). Although MFs enhance tumorigenesis, they are neither mutagenic nor tumorigenic. The mechanism of their tumorigenic effect has not been elucidated. METHODS: To investigate the effect of MFs on apoptosis in HL-60 cells, we exposed the cells to static MFs of 6 mT generated by a magnetic disk of known intensity. Apoptosis was triggered by the DNA topoisomerase I inhibitor, camptothecin (CPT). Activation of caspases in situ using the fluorochrome-labeled inhibitor (FLICA) method and determination of plasma membrane integrity by excluding propidium iodide (PI) were measured by both laser scanning cytometry (LSC) and flow cytometry (FC). LSC and FC identified cells at three sequential stages of their demise: early apoptosis (cells with activated caspases and PI negative); late apoptosis (cells with activated caspases but unable to exclude PI); secondary necrosis (cells with apoptotic morphology no longer stained with FLICA, not excluding PI). RESULTS: MF alone did not induce any apoptogenic or necrogenic effect. CPT exposure led to the sequential appearance of apoptotic cells. In the presence of CPT and MF, the overall proportion of cells undergoing apoptosis was not significantly changed. However, we consistently observed a significant increase in the frequency of late apoptotic/necrotic cells when compared with samples treated with CPT alone (P < 0.001), as well as a decrease in the percentage of early apoptotic cells (P = 0.013). The data obtained by FC and LSC were consistent with each other, showing a similar phenomenon. CONCLUSION: Whereas MF alone or with CPT did not affect overall cell viability, it accelerated the rate of cell transition from apoptosis to secondary necrosis after induction of apoptosis by the DNA-damaging agent, CPT. Modulation of the kinetics of the transition from apoptosis to secondary necrosis by MF in vivo may play a role in inflammation and tumorigenesis.

Apoptosis↗
PubMed2003

Automated red blood cell differential analysis on a multi-angle light scatter/fluorescence hematology analyzer.

BACKGROUND: Most hematology analyzers today are capable of performing white blood cell differential analysis but only limited red blood cell (RBC) differential parameters are available. Because of incomplete morphological information on RBC abnormalities, 5-10% of samples in hematology laboratories routinely undergo smear review. A more complete automated RBC differential capability is desired. METHODS: Abbott CELL DYN 4000 analyzer was modified to perform three-dimensional (3D) RBC differential analysis (RBC/diff) on cell-by-cell basis at 488 nm as well as at 633 nm. Immature RBCs and all nucleated cells are labeled with a fluorescent nuclear stain, prior to RBC/diff using light loss and forward scatter (FSC) signal or two FSC signals and a third side scatter signal, projecting the cytogram onto a precalibrated 3D surface containing grid lines of volume (V) and hemoglobin concentration (HC), to determine the V and HC of a cell. Abnormally shaped RBCs (AbnRBC) are quantitated by the distance of each event to the 3D surface. A total of 154 normal and 484 clinical samples with various hemoglobinopathies have been analyzed. RESULTS: The 3D cytogram and the bivariate distribution of V and HC of individual event provided information to extract quantitative data on V, HC, schistocytes, and AbnRBC of mature and immature RBCs. The accuracy level of the 3D data is difficult to assess because of the semiquantitative nature of smear review, the only available reference method. CONCLUSIONS: The 3D RBC cytograms provide a wealth of morphologic information useful for diagnosis and treatment of patients. With powerful computer programs available today, it is evident that the rate of smear review can be significantly reduced as a result.

Equipment Design↗
PubMed2003

Chemical cytometry.

Chemical cytometry refers to the use of high-sensitivity analytical tools to characterize single cells. These tools include mass spectrometry, electrochemistry and capillary separation methods. This review focuses on the use of capillary electrophoresis coupled with high-sensitivity detection to characterize single cells. In survey experiments, biogenic amines and proteins have been characterized in single cells. In directed experiments, fluorescent substrates are used to monitor the activity of sets of enzymes, either within a family or along an enzymatic cascade. When combined with classical cytometry tools, it is now possible to monitor several cellular components in single cells as a function of cell cycle, which provides insight into the evolution of cellular composition as cells prepare for division.

Cell Cycle↗
PubMedDate not supplied

Iron particle labeling of haematopoietic progenitor cells: an in vitro study.

We present a method for labeling bone marrow haematopoietic progenitor cells with iron particles. Labeling was assessed by magnetic resonance imaging and electron microscopy. Labeling with iron particles could allow the following by imaging techniques of haematopoietic cells in physiologic and pathologic conditions such as the engraftment of haematopoietic progenitor cells or the migration of myelomonocytic cells in inflammatory diseases.

Animals↗
PubMed2000

Analysis of DNA-ploidy using laser scanning cytometer in brain tumors and its clinical application.

There have been few reports on investigation of the cell kinetics of brain tumors using a laser scanning cytometer (LSC). We compared DNA analysis using a flow cytometer (FCM) with that by LSC using established cell lines of brain tumors, and we demonstrated the similarity of both analytical results. The DNA-index (DI) and proliferating index (PI) of various brain tumors were determined, and the results showed that these indices reflected the malignancy of the tumors. The DI values were higher for astrocytoma grade IV than for astrocytoma grade II, frequently showing an aneuploid pattern. The determination of DNA-ploidy using LSC, together with cryopathological diagnosis, was considered to be useful for deciding the area to be resected for glioma perioperatively and to be worth applying clinically.

Brain Neoplasms↗