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Visualisation And Profiling Of Lipids In Single Biological Cells Using Time Of Flight Secondary Ion Mass Spectrometry
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Book Synopsis Visualisation and Profiling of Lipids in Single Biological Cells Using Time-of-Flight Secondary Ion Mass Spectrometry by : Hua Tian
Download or read book Visualisation and Profiling of Lipids in Single Biological Cells Using Time-of-Flight Secondary Ion Mass Spectrometry written by Hua Tian and published by . This book was released on 2011 with total page 155 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Investigating Lipid Heterogeneity in Single Cells Using Time-of-flight Secondary Ion Mass Spectrometry by : Paul D. Piehowski
Download or read book Investigating Lipid Heterogeneity in Single Cells Using Time-of-flight Secondary Ion Mass Spectrometry written by Paul D. Piehowski and published by . This book was released on 2009 with total page 182 pages. Available in PDF, EPUB and Kindle. Book excerpt: Imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) can be utilized to map the spatial distribution of small molecules on a surface with potentially submicron resolution. Due to the inherent characteristics of this technique and its potential to provide higher spatial resolution than light microscopy based techniques without the use of chemical labels, it has been utilized to study the distribution of phospholipid species in the cell membrane. It is now known that many cell membranes contain transient compositional heterogeneities, colloquially referred to as domains, which participate in vital physiological processes such as exocytosis and signal transduction. Because of their size and lifetime, much remains unknown about the nature of these heterogeneities. ToF-SIMS imaging combined with cryogenic sample preparation techniques is a promising analytical platform poised to contribute greatly to this growing field of study. Sample preparation is crucial to obtaining quality lipid distribution maps, especially when dealing with single biological cells. To achieve this end the Winograd and Ewing groups have developed a freeze-fracture methodology adapted from cryo-SEM studies. Freeze-etching, the practice of removing excess surface water from a sample through sublimation into the vacuum of the analysis environment, has also been extensively used in conjunction with electron microscopy. This technique has been applied to ToF-SIMS imaging of cryogenically preserved single cells. By removing the excess water which condenses onto the sample in vacuo, a uniform surface is produced that is ideal for imaging by static SIMS. I demonstrate that the conditions employed to remove deposited water do not adversely affect cell morphology and do not redistribute molecules in the topmost surface layers. In addition, I found that water can be controllably re-deposited onto the sample at temperatures below -100° C in vacuum. The re-deposited water increases the ionization of characteristic fragments of biologically interesting molecules 2-fold without loss of spatial resolution. The utilization of freeze-etch methodology will increase the reliability of cryogenic sample preparations for SIMS analysis by providing greater control of the surface environment. Using these procedures, high quality spectra with both atomic bombardment as well as C60+ cluster ion bombardment, have been obtained. To date, many cell imaging studies have concentrated on phosphatidylcholine distributions, owing to its abundance and high ionization efficiency. However, cholesterol is a particularly interesting molecule due to its involvement in numerous biological processes. For many studies, the effectiveness of chemical mapping is limited by low signal intensity from various bio-molecules. Due to the high energy nature of the SIMS ionization process, many molecules are identified by detection of characteristic fragments. Commonly, fragments of a molecule are identified using standard samples, and those fragments are used to map the location of the molecule. MS/MS data obtained from a prototype C60+/ quadrupole time-of-flight mass spectrometer was used in conjunction with indium LMIG imaging to map previously unrecognized cholesterol fragments in single cells. A model system of J774 macrophages doped with cholesterol was used to show that these fragments are derived from cholesterol in cell imaging experiments. Examination of relative quantification experiments reveals that m/z 147 is the most specific diagnostic fragment and offers a 3-fold signal enhancement. These findings greatly increase the prospects for cholesterol mapping experiments in biological samples, particularly with single cell experiments. In addition, these findings demonstrate the wealth of information that is hidden in the traditional ToF-SIMS spectrum. In order for this technique to provide insight into biological processes, it is critical to characterize the figures of merit. Because a SIMS instrument counts individual events, the precision of the measurement is controlled by counting statistics. As the analysis area decreases, the number of molecules available for analysis diminishes. This becomes critical when imaging sub-cellular features; it limits the information obtainable, resulting in images with only a few counts of interest per pixel. Many features observed in low intensity images are artifacts of counting statistics, making validation of these features crucial to arriving at accurate conclusions. With ToF-SIMS imaging, the experimentally attainable spatial resolution is a function of the molecule of interest, sample matrix, concentration, primary ion, instrument transmission, and spot size of the primary ion beam. A model, based on Poisson statistics, has been developed to validate SIMS imaging data when signal is limited. This model can be used to estimate the effective spatial resolution and limits of detection prior to analysis, making it a powerful tool for tailoring future investigations. In addition, the model allows for pixel-to-pixel intensity comparisons and can be used to validate the significance of observed image features. The implications and capabilities of the model are demonstrated here by imaging the cell membrane of resting RBL-2H3 mast cells. Mass spectrometry imaging has been used to demonstrate that changes in membrane structure drive lipid domain formation in mating single-cell organisms. Chemical studies of lipid bilayers in both living and model systems have revealed that chemical composition is coupled to localized membrane structure. However, it is not clear if the lipids that compose the membrane actively modify membrane structure or if structural changes cause heterogeneity in the surface chemistry of the lipid bilayer. ToF-SIMS images of mating Tetrahymena thermophila, acquired at various stages during mating, can be used to demonstrate that lipid domain formation follows rather than precedes structural changes in the membrane. Domains are formed in response to structural changes that occur during cell-to-cell conjugation. This observation has wide implications in all membrane processes. There is considerable interest in the unique properties of cluster ion projectiles and investigations of how they may be utilized to improve biological imaging. A C60+ cluster ion projectile was employed for sputter cleaning biological surfaces to reveal spatio-chemical information obscured by contamination overlayers. This protocol is used as a supplemental sample preparation method for time of flight secondary ion mass spectrometry (ToF-SIMS) imaging of frozen and freeze dried biological materials. Following the removal of nanometers of material from the surface using sputter cleaning; a frozen-patterned cholesterol film and a freeze-dried tissue sample were analyzed using ToF-SIMS imaging. In both experiments, the chemical information was maintained after the sputter dose, due to the minimal chemical damage caused by C60+ bombardment. The damage to the surface produced by freeze-drying the tissue sample was found to have a greater effect on the loss of cholesterol signal than the sputter-induced damage. In addition to maintaining the chemical information, sputtering is not found to alter the spatial distribution of molecules on the surface. This approach removes artifacts that may obscure the surface chemistry of the sample and are common to many biological sample preparation schemes for ToF-SIMS imaging. In general, out results show that by removing these artifacts, the number of analyzable samples for SIMS imaging is greatly expanded. Although imaging with sub-cellular spatial resolution has been demonstrated, it is clear that the success of future experiments is limited by the ionization efficiency of the lipids, as well as limitations imposed by a coaxial ToF geometry. Considerable work has been done in the lab, to address these limitations. This effort has resulted in the development of a hybrid quadrupole orthogonal ToF instrument equipped with a C60+ primary ion source. The capabilities and potential of this new platform will greatly increase the contributions of SIMS to the biological sciences.
Book Synopsis Development and Application of Methods for Mass Spectrometry Imaging of Lipids Across Biological Surfaces by : Michael Edward Kurczy
Download or read book Development and Application of Methods for Mass Spectrometry Imaging of Lipids Across Biological Surfaces written by Michael Edward Kurczy and published by . This book was released on 2009 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Time of flight secondary ion mass spectrometric (ToF-SIMS) imaging is a powerful bioanalytical tool with the ability to produce molecular images of samples with submicron spatial resolution without the use of labels. In this thesis I will present the development of ToF-SIMS imaging methodology for biological analyses as well as applications that have yielded information about the role of lipids in membrane organization. In the first chapter, I introduce the plasma membrane and describe its fundamental role in maintaining life through the dynamic remodeling of its structure. I focus on two concepts that are believed to influence the localized chemical make up and structure of the membrane, intrinsic curvature and lipid domains. ToF-SIMS imaging is briefly described and a discussion of cluster ion bombardment and sample preparation is included. The chapter concludes with a survey of several important biological studies that have come out of the SIMS community. In Chapter 2 I report a protocol for the use of SIMS imaging to comparatively quantify the relative difference in cholesterol level between the plasma membranes of two cells. This development enables direct comparison of the chemical effects of different drug treatments and incubation conditions in the plasma membrane at the single-cell level. Relative, quantitative ToF-SIMS imaging was used to compare macrophage cells treated to contain elevated levels of cholesterol with respect to control cells. In-situ fluorescence microscopy with two different membrane dyes was used to discriminate morphologically similar but differentially treated cells prior to SIMS analysis. SIMS images of fluorescently identified cells reveal that the two populations of cells have distinct outer leaflet membrane compositions with the membranes of the cholesterol-treated macrophages containing more than twice the amount of cholesterol of control macrophages. Relative quantification with SIMS to compare the chemical composition of single-cells can provide valuable information about normal biological functions, causative agents of diseases, and possible therapies for diseases. Chapter 3 investigates prospects for three-dimensional SIMS analysis of biological materials using model multilayer structures and single cells. The samples were analyzed in a ToF-SIMS spectrometer equipped with a 20 and a 40 keV buckminsterfullerene (C60+) ion source. Specifically, molecular depth profile studies involving dehydrated dipalmitoylphosphatidylcholine (DPPC) organic films indicate that cell membrane lipid materials do not experience significant chemical damage when bombarded with C60+ ion fluences greater than 1015 ions/cm2. Moreover, depth profile analyses of DPPC?sucrose frozen multilayer structures suggest that biomolecule information can be uncovered after the C60+ sputter removal of a 20 nm overlayer with no appreciable loss of underlying molecular signal. The resulting depth information was used to characterize C60+ bombardment of biological materials. This information was used to controllably remove the plasma membrane of a single macrophage cell using a molecular depth profile approach allowing the analysis of the chemistry of the cytoplasm. Two methods that were developed to increase the reproducibility of biological SIMS analysis are covered in Chapter 4. First I demonstrate the utility of the C60+ cluster ion projectile for sputter cleaning biological surfaces to reveal obscured spatio-chemical information. Following the removal of nanometers of material from the surface using sputter cleaning; a frozen-patterned cholesterol film and a freeze-dried tissue sample were analyzed using ToF-SIMS imaging. In both experiments the chemical information was maintained after the sputter dose, due to the minimal chemical damage caused by C60+ bombardment. In fact, the damage to the surface produced by freeze-drying the tissue sample was found to have a greater effect on the loss of cholesterol signal than the sputter-induced damage. In addition to maintaining the chemical information, sputtering did not alter the spatial distribution of the surface chemistry. This approach removes artifacts that are common to many biological sample preparation schemes for ToF-SIMS imaging. Removing these artifacts, which may obscure the surface chemistry of the sample, will increase the number of analyzable samples for SIMS imaging. The second method covered in Chapter 4 is freeze-etching, the practice of removing excess surface water from a sample through sublimation into the vacuum of the analysis environment. This method was used to cryogenically preserve single cells for ToF-SIMS imaging analysis. By removing the excess water, which condenses onto the sample in vacuo, a uniform surface is produced that is ideal for imaging by static SIMS. I demonstrate that the conditions employed to remove deposited water do not adversely affect cell morphology and do not redistribute molecules in the top most surface layers. In addition, I found water could be controllably re-deposited onto the sample at temperatures below -100 oC in vacuum. The re-deposited water increases the ionization of characteristic fragments of biologically interesting molecules 2-fold without loss of spatial resolution. The utilization of freeze-etch methodology will increase the reliability of cryogenic sample preparations for SIMS analysis by providing greater control of the surface environment. Using these procedures we have obtained high quality images and spectra with both atomic bombardment as well as C60+ cluster ion bombardment. Sample handling is also the topic of Chapter 5. It this chapter, I describe a device which has been designed to prepare frozen, hydrated single cell cultures with a freeze fracture methodology for ToF-SIMS analysis in an ION-TOF (GmbH) TOF-SIMS IV mass spectrometer. The device reproducibly produces frozen hydrated sample surfaces for SIMS analysis. I show that SIMS analysis with the Bi32+ produces high-resolution molecular images of single PC12 cells in an ice matrix. I also show that the combination of ionization enhancements that are provided by both the ice matrix and the cluster ion source facilitates the localization of lipid ions that have not been localized in these cells previously. Namely, two fragments of phosphatidlyethanolamine (m/z 124 and m/z 142) and a large fragment of phosphatidylcholine (m/z 224). The ability to localize and measure these ions will increase the number of question that SIMS imaging can be used to answer. In Chapter 6 ToF-SIMS imaging was used to demonstrate that lipid domain formation in mating single-cell organisms is driven by changes in membrane structure. Studies of lipid bilayers in both living and model systems have revealed that lipid composition is coupled to localized membrane structure. However, it is still not clear if the lipids that compose the membrane actively modify membrane structure or if it is structural changes that cause lipid heterogeneity. I report that time of flight secondary ion mass spectrometry images of mating Tetrahymena thermophila acquired before, during and after mating demonstrate that lipid domain formation, identified as a decrease in the lamellar lipid phosphatidylcholine, does not precede structural changes in the membrane. Rather, domains are formed in response to function during cell-to-cell conjugation. ToF-SIMS imaging has been used to collect information with wide implications in all membrane processes. The work presented here is the continuation of a project aimed at chemically characterizing biological samples with spatially resolved mass spectra, with a particular focus on single cell imaging. Much of the work I have done has centered on understanding the capability of current technology and using this understanding to solve a particular problem. This work is vital to keeping SIMS in the biological realm but the development of new technology is the ultimate future for these experiments by increasing the number of tools that the experimenter has to choose from. In Chapter 7 discuss two ongoing projects that I think will lead to the next break through bringing us closer to realizing the goal of this project: a complete chemical map of a single cell.
Book Synopsis Lipidomics and Bioactive Lipids: Mass Spectrometry Based Lipid Analysis by :
Download or read book Lipidomics and Bioactive Lipids: Mass Spectrometry Based Lipid Analysis written by and published by Elsevier. This book was released on 2007-11-26 with total page 432 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume in the well-established Methods in Enzymology series features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Qualitative Analysis and Quantitative Assessment of Changes in Neutral Glycerol Lipid Molecular Species within Cells; Glycerophospholipid identification and quantitation by electrospray ionization mass spectrometry; Detection and Quantitation of Eicosanoids via High Performance Liquid Chromatography/Electrospray Ionization Mass Spectrometry; Structure-specific, quantitative methods for "lipidomic" analysis of sphingolipids by tandem mass spectrometry; Analysis of Ubiquinones, Dolichols and Dolichol Diphosphate-Oligosaccharides by Liquid Chromatography Electrospray Ionization Mass Spectrometry; Extraction and Analysis of Sterols in Biological Matrices by High-Performance Liquid Chromatography Electrospray Ionization Mass Spectrometry; The Lipid Maps Initiative in Lipidomics; Basic analytical systems for lipidomics by mass spectrometry in Japan; The European Lipidomics Initiative Enabling technologies; Lipidomic analysis of Signaling Pathways; Bioinformatics for Lipidomics; Mediator Lipidomics: Search Algorithms for Eicosanoids, Resolvins and Protectins; A guide to biochemical systems modeling of sphingolipids for the biochemist; and Quantitation and Standardization of Lipid Internal Standards for Mass Spectroscopy.
Book Synopsis Mass Spectrometry for Lipidomics by : Michal Holcapek
Download or read book Mass Spectrometry for Lipidomics written by Michal Holcapek and published by John Wiley & Sons. This book was released on 2023-03-07 with total page 897 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass Spectrometry for Lipidomics All-in-one guide to successful lipidomic analysis, combining the latest advances and best practices from academia, industry, and clinical research Mass Spectrometry for Lipidomics presents a systematic overview of lipidomic analysis, covering established standards of lipid analysis, available technology, and key lipid classes, as well as applications in basic research, medicine, pharma, and the food industry. Through connecting recent technological advances with key application areas, this unique guide bridges the gap between academia and industry by translating the vast body of knowledge that has been gained in the past decade into much-needed practical advice for novices as well as routine users. Edited by the president and vice-president of the International Lipidomics Society with contributions from the top experts in lipid analysis, Mass Spectrometry for Lipidomics covers a wide range of key topics, including: Aspects of sample preparation, separation methods, different mass spectrometry modes, as well as identification and quantitation, including the use of bioinformatics tools for data analysis Identification, quantitation and profiling of lipids in different types of biological samples Analytical approaches for all major classes of biological lipids, from fatty acids to phospholipids to sterols Novel applications in biological research, clinical diagnostics, as well as food and crop science For analytical chemists, biochemists, clinical chemists, and analytical laboratories and hospitals, Mass Spectrometry for Lipidomics presents a comprehensive and authoritative overview of the subject, with unmatched expertise from practicing professionals actively involved in the latest research.
Download or read book Lipidomics written by Xianlin Han and published by John Wiley & Sons. This book was released on 2016-04-06 with total page 528 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covers the area of lipidomics from fundamentals and theory to applications Presents a balanced discussion of the fundamentals, theory, experimental methods and applications of lipidomics Covers different characterizations of lipids including Glycerophospholipids; Sphingolipids; Glycerolipids and Glycolipids; and Fatty Acids and Modified Fatty Acids Includes a section on quantification of Lipids in Lipidomics such as sample preparation; factors affecting accurate quantification; and data processing and interpretation Details applications of Lipidomics Tools including for Health and Disease; Plant Lipidomics; and Lipidomics on Cellular Membranes
Book Synopsis Lipidomics and Bioactive Lipids: Lipids and Cell Signaling by :
Download or read book Lipidomics and Bioactive Lipids: Lipids and Cell Signaling written by and published by Elsevier. This book was released on 2007-11-12 with total page 387 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume in the well-established Methods in Enzymology series features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Phospholipase A1 assays using a radio-labeled substrate and mass spectrometry; Real-time Cell Assays of Phospholipases A2 Using Fluorogenic Phospholipids; Analysis and Pharmacological Targeting of Phospholipase C â interactions with G proteins; Biochemical Analysis of Phospholipase D.; Measurement of Autotaxin/Lysophospholipase D Activity; Platelet-Activating Factor; Quantitative measurement of PtdIns(3,4,5)P3; Measuring Phosphorylated Akt And Other Phosphoinositide 3-Kinase-Regulated Phosphoproteins In Primary Lymphocytes; Regulation of Phosphatidylinositol 4-Phosphate 5-Kinase activity by partner proteins; Biochemical Analysis of Inositol Phosphate Kinases; Analysis of the phosphoinositides and their aqueous metabolites; Combination of C17-sphingoid base homologues and mass spectrometry analysis as a new approach to study sphingolipid metabolism; Measurement of mammalian sphingosine-1-phosphate phosphohydrolase activity in vitro and in vivo; A rapid and sensitive method to measure secretion of sphingosine-1-phosphate; Ceramide Kinase and Ceramide-1-Phosphate; Measurement of Mammalian Diacylglycerol Kinase Activity in vitro and in Cells; Lipid Phosphate Phosphatases from Saccharomyces cerevisiae.
Book Synopsis Lipid Characterization with Time-of-flight Secondary Ion Mass Spectrometry (ToF-SIMS) by : Melissa Kathleen Passarelli
Download or read book Lipid Characterization with Time-of-flight Secondary Ion Mass Spectrometry (ToF-SIMS) written by Melissa Kathleen Passarelli and published by . This book was released on 2011 with total page 182 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Lipid Analysis written by W. W. Christie and published by Elsevier. This book was released on 2010-01-10 with total page 447 pages. Available in PDF, EPUB and Kindle. Book excerpt: This well-known and highly successful book was first published in 1973 and has been completely re-written in subsequent editions (published in 1982 and 2003). This new Fourth Edition has become necessary because of the pace of developments in mass spectrometry of intact lipids, which has given recognition of lipid analysis and ‘lipidomics’ as a distinct science. To bring the book up to date with these developments, author William W. Christie is joined by co-author Xianlin Han. Although devoting considerable space to mass spectrometry and lipidomics, Lipid analysis remains a practical guide, in one volume, to the complexities of the analysis of lipids. As in past editions, it is designed to act as a primary source, of value at the laboratory bench rather than residing on a library shelf. Lipid analysis deals with the isolation, separation, identification and structural analysis of glycerolipids, including triacylglycerols, phospholipids, sphingolipids, and the various hydrolysis products of these. The chapters follow a logical sequence from the extraction of lipids to the isolation and characterization of particular lipid classes and of molecular species of each, and to the mass spectrometric analysis of lipids and lipidomics. The new influence of mass spectrometry is due mainly to the development of electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI). Most emphasis in this book is placed on ESI, which is enabling structural characterization of different lipid classes and the identification of novel lipids and their molecular species.
Book Synopsis Lipidomics and Bioactive Lipids: Specialized Analytical Methods and Lipids in Disease by :
Download or read book Lipidomics and Bioactive Lipids: Specialized Analytical Methods and Lipids in Disease written by and published by Elsevier. This book was released on 2007-11-09 with total page 292 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume in the well-established Methods in Enzymology series features methods for the study of lipids using mass spectrometry techniques. Articles in this volume cover topics such as Liquid chromatography mass spectrometry for quantifying plasma lysophospholipids: potential biomarkers for cancer diagnosis; Measurement of eicosanoids in cancer tissues; Noninvasive Assessment of the Role of Cyclooxygenases in Cardiovascular HealthA Detailed HPLC/MS/MS Method; Lipidomics in Diabetes and the Metabolic Syndrome; LC-MS-MS Analysis of Neutral Eicosanoids; Quantification Of F2-Isoprostanes In Biological Fluids And Tissues As A Measure Of Oxidant Stress; Measurement of Products of Docosahexaenoic Acid Peroxidation, Neuroprostanes, and Neurofurans; Enantiomeric separation of hydroxy and hydroperoxy eicosanoids by chiral column chromatography; Targeted Chiral Lipidomics Analysis by Liquid Chromatography Electron Capture Atmospheric Pressure Chemical Ionization Mass Spectrometry (LC-ECAPCI/MS); Shotgun Lipidomics by Tandem Mass Spectrometry under Data-Dependent Acquisition Control; Identification of Intact Lipid Peroxides by Ag+ Coordination Ionspray Mass Spectrometry (CIS-MS); Quantification of Cardiolipin by Liquid Chromatography Electrospray Ionization Mass Spectrometry.
Book Synopsis Life - As a Matter of Fat by : Ole G. Mouritsen
Download or read book Life - As a Matter of Fat written by Ole G. Mouritsen and published by Springer Science & Business Media. This book was released on 2005-10-24 with total page 273 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents a multi-disciplinary perspective on the physics of life and the particular role played by lipids and the lipid-bilayer component of cell membranes. Emphasizes the physical properties of lipid membranes seen as soft and molecularly structured interfaces. By combining and synthesizing insights obtained from a variety of recent studies, an attempt is made to clarify what membrane structure is and how it can be quantitatively described. Shows how biological function mediated by membranes is controlled by lipid membrane structure and organization on length scales ranging from the size of the individual molecule, across molecular assemblies of proteins and lipid domains in the range of nanometers, to the size of whole cells. Applications of lipids in nano-technology and biomedicine are also described.
Book Synopsis Lipidomics in Health & Disease by : Xiangdong Wang
Download or read book Lipidomics in Health & Disease written by Xiangdong Wang and published by Springer. This book was released on 2018-08-30 with total page 204 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume covers the emerging area of science, Clinical Lipidomics, which is the application of lipidology to the understanding of physiological and pathophysiological changes of lipidomes, with a special focus on lipidomic profiles in human diseases. Lipidomics is widely used to map lipid molecular species in a biological system. Clinical lipidomic analysis has demonstrated the comprehensive characterization of molecular lipids in various severities, durations, and therapies as a critical tool in identification and validation of disease-specific biomarkers. This volume on Clinical Lipidomics will add to the literature and help advance the knowledge of the pathogenesis, diagnosis, prevention and treatment of diseases.
Download or read book Lipidomics written by William J Griffiths and published by Royal Society of Chemistry. This book was released on 2020-01-23 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: Lipidomics is one of the emerging ‘omics’ techniques with growing importance in bioscience. Discussing interesting standard and non-standard techniques relevant to the measurement and analysis of lipids by mass spectrometry, this book will provide a guide to the possibilities of the techniques. It will introduce the reader to exciting new methods that allow isomer differentiation, improve sensitivity, allow spatial location and go beyond annotation of simply matching a mass to a database entry. The book is written and edited by the some of the world leaders in the field of lipid mass spectrometry and will have international appeal in industry and academia for analytical chemists, biochemists and biotechnologists. Furthermore, it will provide a useful resource for anyone interested in lipid structure characterization particularly for graduates and postgraduates who require a starting point for their projects.
Book Synopsis Development of Imaging Mass Spectrometry Analysis of Lipids in Biological and Clinically Relevant Applications by : Nathan Heath Patterson
Download or read book Development of Imaging Mass Spectrometry Analysis of Lipids in Biological and Clinically Relevant Applications written by Nathan Heath Patterson and published by . This book was released on 2016 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry is the measurement of the mass over charge ratio of ions. It is broadly applicable and capable of analyzing complex mixtures. Imaging mass spectrometry (IMS) is a branch of mass spectrometry that analyses ions across a surface while conserving their spatial organization on said surface. At this juncture, the most studied IMS samples are thin tissue sections from plants and animals. Among the molecules routinely imaged by IMS, lipids have generated significant interest. Lipids are important in disease and normal cell function as they form cell membranes and act as signaling molecules for cellular events among many other roles. Considering the potential of lipids in biological and clinical applications and the capability of MALDI to ionize lipids, we developed analytical strategies for the handling of samples and analysis of large lipid MALDI IMS datasets. Lipid degradation is massively important in the food industry with oxidized products producing a bad smell and taste. Similarly, lipids in thin tissue sections cut from whole tissues are subject to degradation, and their degradation products can introduce IMS artifacts and the loss of normally occurring species to degradation can skew accuracy in IMS measures of abundance. Oxidized lipids are also known to be important mediators in the progression of several diseases and their accurate preservation is critical. As IMS studies become multi-institutional and collaborations lead to sample exchange, the need for validated protocols and measures of degradation are necessary. We observed the products of lipid degradation in tissue sections from multiple mouse organs and reported on the conditions promoting and inhibiting their presence as well as the timeline of degradation. Our key findings were the increase in oxidized phospholipids and lysophospholipids from degradation at ambient conditions, the decrease in the presence of lipids containing unsaturations on their fatty acyl chains, and the inhibition of degradation by matrix coating and cold storage of sections under N2 atmosphere. At ambient atmospheric and temperature, lipids degraded into oxidized phospholipids on the time-scale of a normal IMS experiment sample preparation (within 30 min). Lipids then degraded into lysophospholipids' on a time scale on the order of several days. Validation of sample handling is especially important when a greater number of samples are to be analyzed either through a cohort of samples, or analysis of multiple sections from a single tissue as in serial 3D IMS. Atherosclerosis is disease caused by accumulation of cellular material at the arterial wall. The accumulation implanted in the cell wall grows and eventually occludes the blood vessel, or causes a stroke. Atherosclerosis is a 3D phenomenon and serial 3D IMS is useful for its ability to localize molecules throughout the length of a plaque and help to define the molecular mechanisms of plaque development and rupture. Serial 3D IMS has many challenges, many of which are simply a matter of producing 3D reconstructions and interpreting them in a timely fashion. In this aim and using analysis of lipids from atherosclerotic plaques from a human carotid and mouse aortic sinuses, we described 3D reconstruction methods using open-source software. Our methodology provides means to obtain high quality visualizations and demonstrates strategies for rapid interpretation of 3D IMS datasets through multivariate segmentation. Mouse aorta from model animals provided a springboard for developing the methods on lower risk samples with less variation with interesting molecular results. 3D MALDI IMS showed localized phospholipid accumulation in the mouse aortic sinuses with correlation between separate positive and negative ionization datasets. Silver-assisted LDI imaging presented differential localization of free fatty acids, cholesterol / cholesterol esters, and triglycerides. The human carotid's 3D segmentation shows molecular histologies (spatial groupings of imaging pixels with similar spectral fingerprints) correlating to the degree of arterial stenosis. Our results outline the potential for 3D IMS in atherosclerotic research. Molecular histologies and their 3D spatial organization, obtained from the IMS techniques used herein, may predict high-risk features, and particularly identify areas of plaque that have higher-risk of rupture. These investigations would help further unravel the biological complexities of atherosclerosis, and predict clinical outcomes. Colorectal cancer liver metastasis (CRCLM) is the metastatic disease of primary colorectal cancer, one of the most common cancers worldwide. CRC is a cancer of the endothelial lining of the colon or rectum. CRC itself is often cured with surgery, while CRCLM is more deadly and treated with chemotherapy with more limited efficacy. Prognosticating and assessment of tumors is performed using classical histopathology with a margin of error. We have used lipid IMS to identify the histological compartments and extract their signatures. Using these IMS signatures we obtained a quantitative and objective histopathological score that correlates with prognosis. Additionally, by dissecting out the lipid signatures we have identified single lipid moieties that are unique to different histologies that could potentially be used as new biomarkers for assessing response to therapy. Particularly, we found a series of plasmalogen and sphingolipid species that differentiate infarct-like and usual necrosis, typical of chemotherapeutic response and normal tumor function, respectively.
Download or read book Lipids written by Gilbert Di Paolo and published by Academic Press. This book was released on 2012-02-16 with total page 503 pages. Available in PDF, EPUB and Kindle. Book excerpt: Lipids are a broad group of naturally occurring molecules which includes fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides, phospholipids, and others. The main biological functions of lipids include energy storage, as structural components of cell membranes, and as important signaling molecules. This volume of Methods in Cell Biology covers such areas as Membrane structure and dynamics, Imaging, and Lipid Protein Interactions. It will be an essential tool for researchers and students alike. Covers such areas as membrane structure and dynamics, imaging, and lipid protein interactions An essential tool for researchers and students alike International authors Renowned editors
Book Synopsis Tandem Mass Spectrometry of Lipids by : Robert C Murphy
Download or read book Tandem Mass Spectrometry of Lipids written by Robert C Murphy and published by Royal Society of Chemistry. This book was released on 2014-12-15 with total page 294 pages. Available in PDF, EPUB and Kindle. Book excerpt: The emerging field of lipidomics has been made possible because of advances in mass spectrometry, and in particular tandem mass spectrometry of lipid ions generated by electrospray ionization. The ability to carry out basic biochemical studies of lipids using electrospray ionization is predicated upon understanding the behaviour of lipid derived ions following collision induced decomposition and mechanisms of product ion formation. During the past 20 years, a wealth of information has been generated about lipid molecules that are now analysed by mass spectrometry, however there is no central source where one can obtain basic information about how these very diverse biomolecules behave following collisional activation. This book will bring together, in one volume, this information so that investigators considering using tandem mass spectrometry to structurally characterize lipids or to quantitate their occurrence in a biological matrix, will have a convenient source to review mechanism of decomposition reactions related to the diversity of lipid structures. A separate chapter is devoted to each of seven major lipid classes including fatty acids, eicosanoids and bioactive lipid mediators, fatty acyl esters and amides, glycerol esters, glycerophospholipids, sphingolipids, and steroids. Mechanistic details are provided for understanding the pathways of formation of major product ions and ions used for structural characterization. In most cases specific ancillary information has been critical to understand the pathways, including isotope labeling and high resolution analysis of precursor and product ions. For a few specific examples such data is missing and pathways are proposed as a means to initiate further mass spectral experiments to prove or disprove pathway hypotheses. While this work largely centres on the lipid biochemistry of animal (mammalian) systems, general principles can be taken from the specific examples and applied to lipid biochemistry found in plants, fungi, prokaryotes and archeal organisms.
Book Synopsis Characterization of Molecular Glycerophospholipids by Quadrupole Time-of-Flight Mass Spectrometry by :
Download or read book Characterization of Molecular Glycerophospholipids by Quadrupole Time-of-Flight Mass Spectrometry written by and published by . This book was released on 2003 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The physical properties of glycerophospholipids (GPLs) are not only determined by the head group (HG), but also by their fatty acid (FA) chains, which affect their distribution and function within membranes in the cell. Understanding the microheterogenity of lipid membranes on a molecular level requires qualitative and quantitative characterization of individual lipids and identification of their FA moieties. The aim of my study was to introduce the new technology of multiple precursor ion scanning (MPIS) on a QSTAR Pulsar time-of-flight mass spectrometer (QqTOF) to analyze lipids. Detailed information on fatty acid composition of individual GPL molecules could be obtained in parallel with conventional profiling of lipid classes, and this could be done by direct analysis of total lipid extracts. This method was termed Fatty Acid Scanning (FAS) and Head Group Scanning HGS, respectively. In this way the molecular GPL composition of total lipid extracts could be charted in a single analysis accurately and rapidly at a low picomole concentration level. Furthermore, combining FAS and HGS together with ion trap MS3 analysis allowed complete charting of the molecular composition of PCs, including quantification of their positional isomers, thus providing a detailed and comprehensive characterization of molecular composition of the pool of PCs. Development of the Lipid Profiler software allowed full automation and rapid processing of complex data, including identification and quantification of molecular GPLs. This approach was evaluated by preliminary applications. First, the molecular composition of PCs of total lipid extracts of MDCK cells and of human red blood cells (RBC) could accurately be charted. Significant presence of positional isomers was observed increasing the total number of individual PC species close to one hundred. Secondly, the molecular PC and SM species distribution in detergent resistant membranes (DRMs) prepared by Triton X-100 DRMs were analyzed and.
