The Experts below are selected from a list of 12381 Experts worldwide ranked by ideXlab platform
Vilmos Kertesz - One of the best experts on this subject based on the ideXlab platform.
-
Laser Capture Microdissection–Liquid Vortex Capture Mass Spectrometry Metabolic Profiling of Single Onion Epidermis and Microalgae Cells
Single Cell Metabolism, 2020Co-Authors: John F. Cahill, Vilmos KerteszAbstract:Laser Capture Microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of Laser Capture Microdissection with mass spectrometry via liquid vortex Capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by Laser Capture Microdissection–liquid vortex Capture mass spectrometry.
-
Laser Capture Microdissection-Liquid Vortex Capture Mass Spectrometry Metabolic Profiling of Single Onion Epidermis and Microalgae Cells.
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: John F. Cahill, Vilmos KerteszAbstract:Laser Capture Microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of Laser Capture Microdissection with mass spectrometry via liquid vortex Capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by Laser Capture Microdissection-liquid vortex Capture mass spectrometry.
Jose Costa - One of the best experts on this subject based on the ideXlab platform.
-
Detection of Ki-ras and p53 Mutations by Laser Capture Microdissection/PCR/SSCP
Methods in molecular biology (Clifton N.J.), 2005Co-Authors: Deborah A. Dillon, Karl Zheng, Brina Negin, Jose CostaAbstract:Efficient detection of somatic mutations is important for the development of clinical molecular diagnostic assays. However, the detection of somatic mutations in tissue is confounded by dilution of the tumor cell population by normal cells. Laser Microdissection allows enrichment for tumor-associated genetic alterations to take place at the level of cell selection, eliminating the need to enrich for mutant alleles after amplification. In this chapter a method is described for somatic mutation analysis using cells acquired by Laser Capture Microdissection.
-
Rapid, efficient genotyping of clinical tumor samples by Laser-Capture Microdissection/PCR/SSCP.
Experimental and molecular pathology, 2001Co-Authors: Deborah A. Dillon, Karl Zheng, Jose CostaAbstract:Background: Mutation analysis is becoming increasingly important in clinical practice, since sporadic mutations in tumors often correlate with prognosis and/or therapeutic response. However, the labor-intensive nature of the molecular analyses has limited the routine clinical use of tumor genotyping. Laser-Capture Microdissection (LCM) allows procurement of relatively pure tumor cell populations. We have investigated the possibility that the use of Laser-Capture Microdissection would allow elimination of time-consuming intermediate steps in tumor genotyping. Design. Archival formalin-fixed, paraffin-embedded tissues from seven cases of colorectal adenocarcinoma were Laser- and hand-microdissected and subsequently evaluated by PCR/SSCP/sequencing for Ki-ras exon 1 and p53 exons 5, 7, and 8. Results. Mutations in Ki-ras exon 1 and/or p53 exons 5 and 7 were detected in five of the seven samples. In the hand-microdissected samples, confident identification of mutations was possible in several cases only after band excision, DNA elution, reamplification, and verification of mutant enrichment by a second SSCP analysis prior to sequencing. In the Laser-microdissected samples, confident mutation identification was possible in all cases with direct sequencing of the original PCR product, reducing the time required for molecular analysis to 3 days. Conclusion. Using Laser-Capture Microdissection, mutant signals are strong enough to sequence directly from original PCR products. With rapid, efficient genotyping by LCM/PCR/SSCP, results can be incorporated directly into the surgical pathology report.
John F. Cahill - One of the best experts on this subject based on the ideXlab platform.
-
Laser Capture Microdissection–Liquid Vortex Capture Mass Spectrometry Metabolic Profiling of Single Onion Epidermis and Microalgae Cells
Single Cell Metabolism, 2020Co-Authors: John F. Cahill, Vilmos KerteszAbstract:Laser Capture Microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of Laser Capture Microdissection with mass spectrometry via liquid vortex Capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by Laser Capture Microdissection–liquid vortex Capture mass spectrometry.
-
Laser Capture Microdissection-Liquid Vortex Capture Mass Spectrometry Metabolic Profiling of Single Onion Epidermis and Microalgae Cells.
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: John F. Cahill, Vilmos KerteszAbstract:Laser Capture Microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of Laser Capture Microdissection with mass spectrometry via liquid vortex Capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by Laser Capture Microdissection-liquid vortex Capture mass spectrometry.
Rosamonde E. Banks - One of the best experts on this subject based on the ideXlab platform.
-
eLS - Laser Capture Microdissection in Proteomics
Encyclopedia of Life Sciences, 2006Co-Authors: Rosamonde E. Banks, Rachel A. Craven, Peter SelbyAbstract:Laser Capture Microdissection or Laser-assisted Microdissection techniques allow the purification or enrichment of selected cell types from heterogeneous tissue sections prior to downstream analysis of DNA, RNA or protein. Keywords: proteomics; Laser Capture Microdissection; Laser-assisted Microdissection; two-dimensional polyacrylamide gel electrophoresis; Western blotting
-
Laser Capture Microdissection for proteome analysis.
Current protocols in protein science, 2003Co-Authors: Rachel A. Craven, Rosamonde E. BanksAbstract:Laser Capture Microdissection is being employed increasingly to isolate specific cell types from tissues, thus overcoming problems of experimental interpretation due to tissue heterogeneity of samples. This unit describes protocols which have been optimized to allow Laser Capture Microdissection of tissues with minimal effect on protein integrity, and their subsequent analysis by techniques including 2D-PAGE, immunoblotting and SELDI, and discusses the relative merits of this approach.
-
Current Protocols in Protein Science - Laser Capture Microdissection for Proteome Analysis
Current Protocols in Protein Science, 2003Co-Authors: Rachel A. Craven, Rosamonde E. BanksAbstract:Laser Capture Microdissection is being employed increasingly to isolate specific cell types from tissues, thus overcoming problems of experimental interpretation due to tissue heterogeneity of samples. This unit describes protocols which have been optimized to allow Laser Capture Microdissection of tissues with minimal effect on protein integrity, and their subsequent analysis by techniques including 2D-PAGE, immunoblotting and SELDI, and discusses the relative merits of this approach.
-
Laser Capture Microdissection and proteomics: possibilities and limitation.
Proteomics, 2001Co-Authors: Rachel A. Craven, Rosamonde E. BanksAbstract:Tissue heterogeneity has always limited the information available from analysis of biological samples in the study of disease. Several approaches have been developed to address this problem, with Laser Capture Microdissection (LCM) emerging as one of the methods of choice. LCM has been extensively used in combination with mutation detection studies and analyses of gene expression at the mRNA level and its potential in proteomics-based research is beginning to be realised. Here we review the progress made to date in the analysis of proteins in LCM-Captured material and evaluate the scope and limitations of this approach.
Lance A. Liotta - One of the best experts on this subject based on the ideXlab platform.
-
Laser Capture Microdissection: Arcturus(XT) infrared Capture and UV cutting methods.
Methods in molecular biology (Clifton N.J.), 2011Co-Authors: Rosa I. Gallagher, Lance A. Liotta, Steven R. Blakely, Virginia EspinaAbstract:Abstract Laser Capture Microdissection (LCM) is a technique that allows the precise procurement of enriched cell populations from a heterogeneous tissue under direct microscopic visualization. LCM can be used to harvest the cells of interest directly or can be used to isolate specific cells by ablating the unwanted cells, resulting in histologically enriched cell populations. The fundamental components of Laser Microdissection technology are (a) visualization of the cells of interest via microscopy, (b) transfer of Laser energy to a thermolabile polymer with either the formation of a polymer-cell composite (Capture method) or transfer of Laser energy via an ultraviolet Laser to photovolatize a region of tissue (cutting method), and (c) removal of cells of interest from the heterogeneous tissue section. Laser energy supplied by LCM instruments can be infrared (810 nm) or ultraviolet (355 nm). Infrared Lasers melt thermolabile polymers for cell Capture, whereas ultraviolet Lasers ablate cells for either removal of unwanted cells or excision of a defined area of cells. LCM technology is applicable to an array of applications including mass spectrometry, DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, proteomics discovery, and signal kinase pathway profiling. This chapter describes the unique features of the Arcturus(XT) Laser Capture Microdissection instrument, which incorporates both infrared Capture and ultraviolet cutting technology in one instrument, using a proteomic downstream assay as a model.
-
Automated Laser Capture Microdissection for tissue proteomics.
Methods in molecular biology (Clifton N.J.), 2008Co-Authors: Adrianna S. Rodriguez, Virginia Espina, Benjamin H. Espina, Lance A. LiottaAbstract:Laser Capture Microdissection (LCM) is a technique for isolating pure cell populations from a heterogeneous tissue section or cytological preparation through direct visualization of the cells. This technique is applicable to molecular profiling of diseased and disease-free tissue, permitting correlation of cellular molecular signatures with specific cell populations. DNA, RNA, or protein analysis may be performed with the microdissected tissue by any method with adequate sensitivity.Automated LCM platforms combine graphical user interfaces and annotation software for visualization of the tissue of interest in addition to robotically controlled Microdissection. The principal components of LCM technology are (1) visualization of the cells of interest through microscopy, (2) transfer of Laser energy to a thermolabile polymer with formation of a polymer-cell composite, and (3) removal of the cells of interest from the heterogeneous tissue section. Automated LCM is compatible with a variety of tissue types, cellular staining methods, and tissue preservation protocols allowing Microdissection of fresh or archival specimens in a high-throughput manner. This protocol describes Microdissection techniques compatible with downstream proteomic analyses.
-
Molecular analysis of microdissected tissue: Laser Capture Microdissection
PCR Applications, 2007Co-Authors: Nicole L. Simone, Rodrigo F. Chuaqui, Lance A. Liotta, Jeffrey Y. Lee, Mary Huckabee, Kristina A. Cole, Chetan Seshadri, Bob Bonner, Michael R. Emmert-buckAbstract:Publisher Summary At the microscopic level, tissues are composed of complicated interacting and interdependent cell populations, which are regulated by the local extracellular matrix. Thus, analyzing critical gene expression patterns in development, normal function, and disease progression depends on the extraction of specific cells from their complex tissue milieu. Laser Capture Microdissection (LCM) is developed to provide a rapid, reliable method to procure pure populations of selected cells from specific microscopic regions of tissue sections for molecular analysis. This chapter discusses the molecular analysis of microdissected tissue. Further, it describes protocols for preparing the tissue before Microdissection and conducting the PCR amplification after Microdissection. Finally, it outlines the applications. DNA obtained from cells procured by LCM from clinical specimens is analyzed by a variety of methods, including loss of heterozygosity, clonal analysis, and direct sequencing. Similarly, LCM is used to study expression differences in various tissues using RNA-based techniques, such as cDNA library construction, microarray hybridization, and differential gene expression.
-
Laser Capture Microdissection technology.
Expert review of molecular diagnostics, 2007Co-Authors: Virginia Espina, Michael Heiby, Mariaelena Pierobon, Lance A. LiottaAbstract:Deciphering the cellular and molecular interactions that drive disease within the tissue microenvironment holds promise for discovering drug targets of the future. In order to recapitulate the in vivo interactions through molecular analysis, one must be able to analyze specific cell populations within the context of their heterogeneous tissue microecology. Laser Capture Microdissection is a method to procure subpopulations of tissue cells under direct microscopic visualization. Laser Capture Microdissection technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, mass spectrometry proteomics discovery and signal pathway profiling.
-
Laser-Capture Microdissection
Nature Protocols, 2006Co-Authors: Virginia Espina, Julia D. Wulfkuhle, Amy Vanmeter, David H. Geho, Emanuel F Petricoin, George Coukos, Valerie S Calvert, Weidong Zhou, Lance A. LiottaAbstract:Deciphering the cellular and molecular interactions that drive disease within the tissue microenvironment holds promise for discovering drug targets of the future. In order to recapitulate the in vivo interactions thorough molecular analysis, one must be able to analyze specific cell populations within the context of their heterogeneous tissue microecology. Laser-Capture Microdissection (LCM) is a method to procure subpopulations of tissue cells under direct microscopic visualization. LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, RNA transcript profiling, cDNA library generation, proteomics discovery and signal-pathway profiling. Herein we provide a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1-1.5 h.
