The Experts below are selected from a list of 113829 Experts worldwide ranked by ideXlab platform
James R. Foreman - One of the best experts on this subject based on the ideXlab platform.
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Five-color immunophenotyping plus DNA Content analysis by laser scanning cytometry
Communications in Clinical Cytometry, 1998Co-Authors: Richard J. Clatch, James R. ForemanAbstract:Laser scanning cytometry is a new laboratory technology similar to flow cytometry but with advantages for certain clinical and research applications. To date, laser scanning cytometry has been successfully used to perform three-color immunophenotypic analysis of hematologic specimens, single-color immunophenotyping plus DNA Content analysis of numerous specimen types, and automated analysis of fluorescence in situ hybridization specimens. Several other interesting applications are also in development. In general, advantages of laser scanning cytometry include reduced specimen size requirements, simplified methodologies, and the ability to microscopically examine individual cells-allowing for the direct correlation of cytologic morphology with objective fluorescence measurements. In this report, we describe a method which more fully takes advantage of the laser scanning cytometer's capabilities for immunophenotypic analysis of hematologic specimens. Specifically, we have devised a method to increase the number of fluorescent parameters from three to a total of six, five representing binding of immunofluorescent antibodies and one for stoichiometric measurements of DNA Content. As with most laser scanning cytometric applications, this technique can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology
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multiparameter analysis of DNA Content and cytokeratin expression in breast carcinoma by laser scanning cytometry
Archives of Pathology & Laboratory Medicine, 1997Co-Authors: Richard J. Clatch, James R. Foreman, Jami L Walloch, Louis A KamentskyAbstract:Objective The objective of this study was to test a new laboratory technology, laser scanning cytometry, for the purpose of performing multiparameter DNA Content analysis of breast carcinomas. Design We developed a simplified method of multiparameter DNA Content analysis using cytokeratin expression to positively gate epithelial cells. Over 300 consecutive cases of breast carcinoma were analyzed by multiparameter laser scanning cytometry. The first 73 cases were analyzed in parallel by single parameter flow cytometry. Setting The Department of Pathology, Christ Hospital and Medical Center, Oak Lawn, Ill. Specimens Three hundred eighteen consecutive cases of breast carcinoma presenting between March 1994 and December 1995. Main outcome measures Outcome measures included the percentage of cases for which DNA Content analysis could be successfully performed given the limitations of specimen size. Additionally, for the first 73 cases, laser scanning cytometry results were compared with flow cytometry results. Results All of the first 73 cases were successfully analyzed by laser scanning cytometry, but for 8 cases (11%) there was insufficient material for flow cytometry. Correlation of DNA Content for the remaining 65 cases analyzed in parallel by the two methods was nearly perfect (p = .994). Five seemingly discrepant cases highlighted the importance of cytokeratin gating of epithelial cells by any technique, as well as other advantages specific to laser scanning cytometry, such as the ability to examine individual cells microscopically and correlate cytologic morphology with DNA Content results. Conclusions Laser scanning cytometry is a promising new technology for DNA Content analysis of solid tissue tumors. Further work needs to be performed to validate the prognostic potential of the laser scanning cytometric assay results and to generate methodologies aimed at providing highly objective determinations of tumor cell S-phase fraction.
Richard J. Clatch - One of the best experts on this subject based on the ideXlab platform.
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Five-color immunophenotyping plus DNA Content analysis by laser scanning cytometry
Communications in Clinical Cytometry, 1998Co-Authors: Richard J. Clatch, James R. ForemanAbstract:Laser scanning cytometry is a new laboratory technology similar to flow cytometry but with advantages for certain clinical and research applications. To date, laser scanning cytometry has been successfully used to perform three-color immunophenotypic analysis of hematologic specimens, single-color immunophenotyping plus DNA Content analysis of numerous specimen types, and automated analysis of fluorescence in situ hybridization specimens. Several other interesting applications are also in development. In general, advantages of laser scanning cytometry include reduced specimen size requirements, simplified methodologies, and the ability to microscopically examine individual cells-allowing for the direct correlation of cytologic morphology with objective fluorescence measurements. In this report, we describe a method which more fully takes advantage of the laser scanning cytometer's capabilities for immunophenotypic analysis of hematologic specimens. Specifically, we have devised a method to increase the number of fluorescent parameters from three to a total of six, five representing binding of immunofluorescent antibodies and one for stoichiometric measurements of DNA Content. As with most laser scanning cytometric applications, this technique can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology
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multiparameter analysis of DNA Content and cytokeratin expression in breast carcinoma by laser scanning cytometry
Archives of Pathology & Laboratory Medicine, 1997Co-Authors: Richard J. Clatch, James R. Foreman, Jami L Walloch, Louis A KamentskyAbstract:Objective The objective of this study was to test a new laboratory technology, laser scanning cytometry, for the purpose of performing multiparameter DNA Content analysis of breast carcinomas. Design We developed a simplified method of multiparameter DNA Content analysis using cytokeratin expression to positively gate epithelial cells. Over 300 consecutive cases of breast carcinoma were analyzed by multiparameter laser scanning cytometry. The first 73 cases were analyzed in parallel by single parameter flow cytometry. Setting The Department of Pathology, Christ Hospital and Medical Center, Oak Lawn, Ill. Specimens Three hundred eighteen consecutive cases of breast carcinoma presenting between March 1994 and December 1995. Main outcome measures Outcome measures included the percentage of cases for which DNA Content analysis could be successfully performed given the limitations of specimen size. Additionally, for the first 73 cases, laser scanning cytometry results were compared with flow cytometry results. Results All of the first 73 cases were successfully analyzed by laser scanning cytometry, but for 8 cases (11%) there was insufficient material for flow cytometry. Correlation of DNA Content for the remaining 65 cases analyzed in parallel by the two methods was nearly perfect (p = .994). Five seemingly discrepant cases highlighted the importance of cytokeratin gating of epithelial cells by any technique, as well as other advantages specific to laser scanning cytometry, such as the ability to examine individual cells microscopically and correlate cytologic morphology with DNA Content results. Conclusions Laser scanning cytometry is a promising new technology for DNA Content analysis of solid tissue tumors. Further work needs to be performed to validate the prognostic potential of the laser scanning cytometric assay results and to generate methodologies aimed at providing highly objective determinations of tumor cell S-phase fraction.
Miroslav Kolařik - One of the best experts on this subject based on the ideXlab platform.
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application of flow cytometry for exploring the evolution of geosmithia fungi living in association with bark beetles the role of conidial DNA Content
Fungal Ecology, 2015Co-Authors: Tereza Veselska, Miroslav KolařikAbstract:Abstract Geosmithia belongs among fungi living in symbiosis with phloem-feeding bark beetles. Several species have altered their ecology to that of obligatory symbiosis with ambrosia beetles, which has led to a shift in their phenotype and caused formation of large spherical conidia. In this study, we pose the following questions; (1) Is the conidial DNA Content of Geosmithia correlated with conidial volume?; (2) Is the DNA Content of Geosmithia related to the degree of mutual dependence between Geosmithia and their vector? There was a positive and strong correlation between conidial DNA Content and conidial volume in Geosmithia . Also species more narrowly associated with the vector tend to have a larger conidial DNA Content and volume than less narrowly associated species. Ambrosia fungi achieved the biggest conidial DNA Content and volume compared to other species. We suppose that polyploidisation occurred during the evolution of ambrosia species in the genus Geosmithia .
Louis A Kamentsky - One of the best experts on this subject based on the ideXlab platform.
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multiparameter analysis of DNA Content and cytokeratin expression in breast carcinoma by laser scanning cytometry
Archives of Pathology & Laboratory Medicine, 1997Co-Authors: Richard J. Clatch, James R. Foreman, Jami L Walloch, Louis A KamentskyAbstract:Objective The objective of this study was to test a new laboratory technology, laser scanning cytometry, for the purpose of performing multiparameter DNA Content analysis of breast carcinomas. Design We developed a simplified method of multiparameter DNA Content analysis using cytokeratin expression to positively gate epithelial cells. Over 300 consecutive cases of breast carcinoma were analyzed by multiparameter laser scanning cytometry. The first 73 cases were analyzed in parallel by single parameter flow cytometry. Setting The Department of Pathology, Christ Hospital and Medical Center, Oak Lawn, Ill. Specimens Three hundred eighteen consecutive cases of breast carcinoma presenting between March 1994 and December 1995. Main outcome measures Outcome measures included the percentage of cases for which DNA Content analysis could be successfully performed given the limitations of specimen size. Additionally, for the first 73 cases, laser scanning cytometry results were compared with flow cytometry results. Results All of the first 73 cases were successfully analyzed by laser scanning cytometry, but for 8 cases (11%) there was insufficient material for flow cytometry. Correlation of DNA Content for the remaining 65 cases analyzed in parallel by the two methods was nearly perfect (p = .994). Five seemingly discrepant cases highlighted the importance of cytokeratin gating of epithelial cells by any technique, as well as other advantages specific to laser scanning cytometry, such as the ability to examine individual cells microscopically and correlate cytologic morphology with DNA Content results. Conclusions Laser scanning cytometry is a promising new technology for DNA Content analysis of solid tissue tumors. Further work needs to be performed to validate the prognostic potential of the laser scanning cytometric assay results and to generate methodologies aimed at providing highly objective determinations of tumor cell S-phase fraction.
Hong Zhao - One of the best experts on this subject based on the ideXlab platform.
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analysis of cellular DNA Content by flow and laser scanning cytometry
Advances in Experimental Medicine and Biology, 2010Co-Authors: Dorota H Halicka, Hong ZhaoAbstract:This chapter covers several aspects of methodology of DNA Content analysis in individual cells that is most commonly used for assessment of DNA ploidy and for enumeration of cells in particular phases of the cell cycle. Briefly presented are general principles of instrumentation and cell analysis by flow- and laser scanning- cytometry. Described are major methods designed to stain DNA with fluorochromes in live cells, in detergent-permeabilized cells, in cells fixed prior to DNA staining as well as in nuclei of cells isolated from paraffin-embedded tissues. Briefly addressed are approaches to estimate cellular DNA Content in conjunction with cellular immunophenotype. Discussed are factors that affect accuracy of DNA Content measurement such as: (i) differences in chromatin structure of the analyzed cells that restrict DNA accessibility to fluorochromes, (ii) stoichiometry of interaction between fluorochromes and DNA in chromatin and (iii) chemical mass action law defining dependency of fluorochrome binding to DNA in relation to fluorochrome concentration and number of potential binding sites in a sample. Described also are controls used to ensure accuracy of DNA ploidy determination, the principles in ploidy assessment and possible pitfalls in analysis.