Tissue Profiling by MALDI-Tof MS: Basic Principles From Small Molecules to Proteins
|
|
- Elwin Hudson
- 6 years ago
- Views:
Transcription
1 Tissue Profiling by MALDI-Tof MS: Basic Principles From Small Molecules to Proteins James Mobley, Ph.D. Proteomics Facility Vanderbilt University Imaging Mass Spectrometry of Thin Tissue Sections To obtain information on the local protein composition at any coordinate on the section To reconstruct -dimensional density maps (or images) for all of the signals detected To obtain molecular profiles and images indicative of health status Discovery tool to answer fundamental questions relevant to protein expression in normal and diseased tissues
2 Slice frozen tissue on cryostat (~ μm thick) Profiling Thaw slice onto MALDI plate, allow to dry Imaging Droplets Apply matrix Spray coating Laser Acquire mass spectra Laser Protein profiles Protein images m/z 8 88 MALDI Time-of-Flight Mass Spectrometry Matrix Sample +/- U Ionizing Probe (start) M M M Ion detector (MCP) Ion signals % Intensity ~ ~
3 Protein expression profiling by MALDI-MS Human breast tumor needle biopsy mm % Intensity x α+ 79 β ~ α + β Intensity Intensity
4 Human glioma biopsy mm C X 5 Tumor A B D Non-Tumor Tumor (A) Tumor (A) Tumor (B) Tumor (B) Non Tumor (C) Non Tumor (C) Non Tumor (D) Non Tumor (D) 8 Schwartz et Al. Clin. Cancer Res.,, (). m/z 5 mm Brain glioma section stained with Methylene blue 5 % Intensity x % Intensity
5 Principle of MALDI MS Imaging Frozen section MS Images Matrix application Raster of section 6 8 m/z MALDI Mass Spectrum Spray deposition of matrix on tissue sections Spray nebulizer for TLC plates Matrix solution Tissue section Nitrogen 5
6 Comparing matrix coatings: Applied Biosystems DE-STR Spotted % Intensity um Laser spot μm Sprayed % Intensity Mouse Brain , c) 759, d) Imaging resolution: 5 μm mm % m/z 67 % Intensity , e) 8, f) x ~99 m/z 759 m/z , g) m/z 8 m/z 8 Chaurand et Al. Anal Chem, 76, 86A-9A (). 6
7 Glioma mouse model - intracranial injection of GL6 cancer cells 965 Tumor mm Tumor Non tumor Relative intensities NT) T) , α , β , α + 568, β x NT) 6 7 T) Glioma mouse model. Imaging resolution: μm mm m/z 69 m/z 759 m/z 99 Acyl CoAbinding protein m/z 7 and 8 Histone H m/z Cytochrome C m/z 8 and ~5 5 Histones HB and H m/z 9 m/z 98 and 5 67 α and β hemoglobins m/z 8 Chaurand et Al. Anal Chem, 76, 86A-9A (). m/z 7 m/z 87 % 7
8 Acoustic Drop Ejection technology provides a nozzle-free means of generating microdroplets on-demand x y Sample plate Ejection of microdroplets Matrix reservoir Coupling membrane Coupling reservoir Transducer RF generator Waveform generator Current performances: - Spot size: ~8- µm - Drop ejection rate: Hz - Drops per pixel: 6-8 Manual spotting vs The RapidSpotter A) Mouse brain, Analysis of the corpus callosum B) 8
9 RapidSpotter Imaging of a Mouse Brain Section m/z 7885 m/z 89 m/z 688 μm Matrix printing % % m/z 79 m/z 6759 m/z 78 Create custom plate file Intensity
10 Schematic Representation of Protein Marker Identification Homogenate Lung Tumor * m/z RP HPLC Fractions Tryptic Digestion MS/MS MALDI MS * 5 5 m/z Mw Matches Mw Matches Add Post- Translational Modification Database Search Protein ID m/z mucosa submucosa Normal human colon biopsy µm muscle m/z 55 Combined Image m/z 79 m/z 8958 Combined Image
11 cortex medula Rat kidney sagittal section m/z 9979 m/z,95.5 mm Intensity m/z 99 m/z 7, Overlay Overlay Molecular Determination of Tumor Margins Histological tumor margin Non-tumor cm 5 x spots 5 µm resolution Tumor Vessel Fibrous stroma Skeletal muscle Robert Caldwell,
12 B A B A C D E E C D Soft Tissue Sarcoma Myosin light chain Histone H isoform (m/z, 7) Histone H isoform (m/z,85) Rob Caldwell
13
14 Histological vs. molecular assessment of the tumor margin: Brain Cancer protein expression profiling by MALDI-MS Laser ; Matrix droplets are regionally deposited on the sample. Each spot area is analyzed using MALDI-MS ; Computer algorithms are used to smooth and correct baseline of each spectrum N N N N T T T T T T T ; Hierarchical cluster analysis of human brain cancer and normal brain spectra Schwartz SA et Al, Cancer Research, 65(7), (5)
15 Weighted Flexible Compound Covariate Method Pairwise comparisons are performed between groups Fisher s exact test Individual protein signals differentially expressed are selected using 6 independent statistical analysis Significance Analysis of Microarrays Kruskal- Wallis test Proteins found in at least analysis were included in the final marker list A pathology prediction model is generated using the top significant protein classifiers Jason Moore, Yu Shyr Vanderbilt University t-test Weighted Gene Analysis Information- Score Method Human glioma protein expression profiling by MALDI-MS Non-Tumor Absolute Intensity Grade Tumor Grade Tumor Grade Tumor 8 5
16 6 % % proteins Grade, and Non-Tumor Clustering analysis of proteomic patterns in human glioma Non-Tumor vs. All Tumor - Training cohort Non-tumor Grade Clustering analysis of proteomic patterns in human glioma Non-Tumor vs. Grade - Training cohort proteins
17 Kaplan-Meier survival curves for groups with poor and good prognostic. Classification according to the expression pattern of 9 distinct MS peaks Long-term survival group (n = 6) % Surviving Patient Population 8 6 P-value <. 9 proteins were significant N = 6 Grade Glioma N = 5 5 Short-term survival group (n = 5) P-value <.5 8 proteins where significant 5 6 Month post surgery Schwartz SA et Al, Cancer Research, 65(7), (5) Human brain glioma Low grade vs. high grade m/z 86, S Beta Relative Intensities HG LG 68 % m/z 86 HG LG Immunohistochemistry HG LG µm 7
18 Analysis of drugs in tissue by mass spectrometry Cut frozen slice ( μm) Dose animal orally i.v. Remove tissue Apply matrix Reyzer ML et al, J Mass Spectrom, 8, 8-9 () Reyzer ML et al, Cancer Res 6, 99-9 () Laser Analyze by: MALDI MS and MALDI MS/MS MMTV/HER transgenic mouse mammary tumors MMTV/HER cells transplanted in FVB female mice. Tumor grown to a size of ~ mm. OSI 77 is an intracellular tyrosine kinase EGF receptor inhibitor. Administered orally for week Tumor Volume (mm ) 8 6 Captisol µl (vector) OSI 77 mg/kg Days post-treatment Contributed by M. Sliwkowski (Genentech, Inc.) 8
19 MS/MS analysis of OSI-77 in tumor tissue Tumors removed after a single mg/kg dose of OSI-77 O O m/z 6. +H O N N O NH +H m/z 78. +H + MW = 9. Intensity 78. [OSI77 + H] + 6. hr HC C 5 Monitored CAD transition m/z Intensity 5 m/z 78 Spot # 6 8 m/z Dose dependence of protein alteration ( hr after dose) 9 control mice, Av of 5 spectra thymosin β x dosed mice, Av of 9 spectra per dose Relative intensity Control mg/kg mg/kg mg/kg
20 Using Protein Fingerprints to Evaluate Drug Therapy drug treatment surgical/ MRI assessment of tumor volume 5 days MS profiling of proteome changes Whole Rat Imaging Sheerin Khatib-Shahid Average Ion Spectrum for Whole Rat Sagittal Section Muscle Thymus Kidney Brain Caudal Liver Cecum Heart m/z wall region Cortex m/z m/z m/z cm
21 Whole Rat Sagittal Section Imaging Rat Head Subsection % 5% cm % signal intensity Optical image Brain White matter Corpus Callosum Eye socket m/z 76 m/z 7 m/z 5978 m/z 6858 Drug Imaging Olanzapine (MW ) N N N CH 8 56 Intensity N H S CH Spleen Kidney Stomach Lung Liver Cecum Heart cm Optical image of orally dosed (8 mg/ml) rat abdomen MALDI MS/MS Image of OLZ (m/z 56) in stomach hours post-dose MS/MS Image of OLZ, x signal intensity with stomach signals masked
22 h Optical Image fur kidney liver lung spinal cord brain testis bladder? thoracic cavity Olanzapine m/z 56 thymus N-desmethyl metabolite m/z hydroxymethyl metabolite m/z 9 7 Areas of Further Development - Increase number of proteins analyzed to >> - Develop sample protocols to investigate peptides - Achieve high sensitivity above 5 kda - Integrate detergents into tissue protocol for membrane associated proteins - Pixel-to-pixel cycle time of about ms needed - Integrate bioinformatic tools, image processing - Develop sample protocols targeted at specific tissues/diseases - Improve protein identification protocols
23 Tissue Profiling/Imaging MS Conclusions - Excellent discovery tool - MW annotated patterns keyed to tissue location - Augments (not replaces) current molecular technologies - Significant clinical potential (disease diagnosis, state and progression, prognosis, risk assessment (?) - Provides temporal proteomic profile that, with genomic profile, will be vital to personalized medicine MSRC Richard Caprioli (Director) Shannon Cornett James Mobley Michelle Reyzer Jeremy Norris Robert Caldwell Erin Seeley Stacey Oppenheimer Sheerin Khatib-Shahid Kristin Burnum Kristen Herring Saturday Puolitaival Hans Reudi Aerni Annette Erskine Ken Shriver Others Per Andren, Uppsala University Robert Weil, NIH Walter Korfmacher, Schering-Plough Markus Stoeckli, Novartis Acknowledgments Vanderbilt Collaborators Carlos Arteaga David Carbone Robert Coffey Jr. Marie-Claire Orgebin-Crist Ariel Deutch Dennis Hallahan Pat Levitt Robert Matusik Jason Moore Hal Moses Jennifer Pietenpol Ned Porter Yu Shyr Robert Whitehead Ken Schriver Funding NIH (NCI, GMS, NIDA) Vanderbilt University
Profiling and Imaging Mass Spectrometry
Profiling and Imaging Mass Spectrometry Compilation of Work from the Caprioli Lab James Mobley, Ph.D. Director of Urologic Research Associate Director of Mass Spectrometry University of Alabama at Birmingham
More informationSequence Identification And Spatial Distribution of Rat Brain Tryptic Peptides Using MALDI Mass Spectrometric Imaging
Sequence Identification And Spatial Distribution of Rat Brain Tryptic Peptides Using MALDI Mass Spectrometric Imaging AB SCIEX MALDI TOF/TOF* Systems Patrick Pribil AB SCIEX, Canada MALDI mass spectrometric
More informationThree Dimensional Mapping and Imaging of Neuropeptides and Lipids in Crustacean Brain
Three Dimensional Mapping and Imaging of Neuropeptides and Lipids in Crustacean Brain Using the 4800 MALDI TOF/TOF Analyzer Ruibing Chen and Lingjun Li School of Pharmacy and Department of Chemistry, University
More informationSmall Molecule Drug Imaging of Mouse Tissue by MALDI-TOF/TOF Mass Spectrometry and FTMS
Bruker Daltonics Application Note # MT-93/FTMS-38 Small Molecule Drug Imaging of Mouse Tissue by MALDI-TOF/TOF Mass Spectrometry and FTMS Introduction Matrix Assisted Laser Desorption Ionization (MALDI)
More informationFuture Directions: Tissue and Cell Imaging Robert C. Murphy
LIPID MAPS Lipidomics Workshop April 19, 2009 www.lipidmaps.org m/z 806 (16:0a/22:6-PC) [M+H] + Future Directions: Tissue and Cell Imaging Robert C. Murphy 16:0/22:6 PC m/z 806.4 Department of Pharmacology
More informationProteomic Biomarker Discovery in Breast Cancer
Proteomic Biomarker Discovery in Breast Cancer Rob Baxter Laboratory for Cellular and Diagnostic Proteomics Kolling Institute, University of Sydney, Royal North Shore Hospital robert.baxter@sydney.edu.au
More informationMALDI-IMS (MATRIX-ASSISTED LASER DESORPTION/IONIZATION
MALDI-IMS (MATRIX-ASSISTED LASER DESORPTION/IONIZATION IMAGING MASS SPECTROMETER) IN TISSUE STUDY YANXIAN CHEN MARCH 8 TH, WEDNESDAY. SEMINAR FOCUSING ON What is MALDI imaging mass spectrometer? How does
More informationMALDI Imaging Mass Spectrometry
MALDI Imaging Mass Spectrometry Nan Kleinholz Mass Spectrometry and Proteomics Facility The Ohio State University Mass Spectrometry and Proteomics Workshop What is MALDI Imaging? MALDI: Matrix Assisted
More informationTechnical Note # TN-31 Redefining MALDI-TOF/TOF Performance
Bruker Daltonics Technical Note # TN-31 Redefining MALDI-TOF/TOF Performance The new ultraflextreme exceeds all current expectations of MALDI-TOF/TOF technology: A proprietary khz smartbeam-ii TM MALDI
More informationSupporting information for: Memory Efficient. Principal Component Analysis for the Dimensionality. Reduction of Large Mass Spectrometry Imaging
Supporting information for: Memory Efficient Principal Component Analysis for the Dimensionality Reduction of Large Mass Spectrometry Imaging Datasets Alan M. Race,,, Rory T. Steven,, Andrew D. Palmer,,,
More informationMass spectrometry imaging. What does MS imaging offer?
BMG 744 Mass spectrometry imaging Stephen Barnes, PhD With sincere acknowledgments to David Stella, PhD and Kyle A. Floyd, MS, former students in the Barnes Laboratory (2005 2012) and Kevin Schey, PhD,
More informationMS-IMS (MALDI-IMAGING)?
MS-IMS (MALDI-IMAGING)? Protein Chemistry/Proteomics and Peptide Synthesis and Array Unit Biomedicum Helsinki and Haartman Institute E-Mail: marc.baumann@helsinki.fi (http://research.med.helsinki.fi/corefacilities/proteinchem)
More informationMass Spectrometry. Mass spectrometer MALDI-TOF ESI/MS/MS. Basic components. Ionization source Mass analyzer Detector
Mass Spectrometry MALDI-TOF ESI/MS/MS Mass spectrometer Basic components Ionization source Mass analyzer Detector 1 Principles of Mass Spectrometry Proteins are separated by mass to charge ratio (limit
More informationBildgebende Massenspektrometrie Erfolgreiche Anwendungen in der biomedizinischen Forschung. Markus Stoeckli
Bildgebende Massenspektrometrie Erfolgreiche Anwendungen in der biomedizinischen Forschung Markus Stoeckli MALDI Mass Spectrometric Imaging Principle 2 MALDI MSI Markus Stoeckli September 2014 Comparison
More informationMALDI Imaging Drug Imaging Detlev Suckau Head of R&D MALDI Bruker Daltonik GmbH. December 19,
MALDI Imaging Drug Imaging Detlev Suckau Head of R&D MALDI Bruker Daltonik GmbH December 19, 2014 1 The principle of MALDI imaging Spatially resolved mass spectra are recorded Each mass signal represents
More informationMALDI-TOF Mass Spectrometry: A New Rapid ID Method in Clinical Microbiology
MALDI-TOF Mass Spectrometry: A New Rapid ID Method in Clinical Microbiology Patrick R. Murray, PhD WW Director, Scientific Affairs BD Diagnostic Systems Outline MALDI-TOF is the most important innovation
More informationIsolation of pure cell populations from healthy and
Direct Analysis of Laser Capture Microdissected Cells by MALDI Mass Spectrometry Baogang J. Xu and Richard M. Caprioli Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA Melinda
More informationMass spectrometry Technologies in Lipid chemistry
Mass spectrometry Technologies in Lipid chemistry Rabah Soliymani University Of Helsinki Protein Chemistry Unit Biomedicum Helsinki Rabah.soliymani@helsinki.fi Complex_&_dynamic_mixtures (few copies to
More informationTissue and Fluid Proteomics Chao-Cheng (Sam) Wang
Tissue and Fluid Proteomics Chao-Cheng (Sam) Wang UAB-03/09/2004 What is proteomics? A snap shot of the protein pattern!! What can proteomics do? To provide information on functional networks and/or involvement
More informationImaging of lipid species by MALDI mass spectrometry
Imaging of lipid species by MALDI mass spectrometry Robert C. Murphy, 1 Joseph A. Hankin, and Robert M. Barkley Department of Pharmacology, MS 8303 University of Colorado Denver, Aurora, CO 80045 Abstract
More informationAnalysis of Peptides via Capillary HPLC and Fraction Collection Directly onto a MALDI Plate for Off-line Analysis by MALDI-TOF
Analysis of Peptides via Capillary HPLC and Fraction Collection Directly onto a MALDI Plate for Off-line Analysis by MALDI-TOF Application Note 219 Joan Stevens, PhD; Luke Roenneburg; Kevin Fawcett (Gilson,
More informationBruker Daltonics. autoflex III smartbeam. The Standard in MALDI-TOF Performance MALDI-TOF/TOF. think forward
Bruker Daltonics autoflex III smartbeam The Standard in MALDI-TOF Performance think forward MALDI-TOF/TOF Designed for a Routine High Level of Performance The autoflex III smartbeam brings the power of
More informationMALDI-TOF. Introduction. Schematic and Theory of MALDI
MALDI-TOF Proteins and peptides have been characterized by high pressure liquid chromatography (HPLC) or SDS PAGE by generating peptide maps. These peptide maps have been used as fingerprints of protein
More informationPTM Discovery Method for Automated Identification and Sequencing of Phosphopeptides Using the Q TRAP LC/MS/MS System
Application Note LC/MS PTM Discovery Method for Automated Identification and Sequencing of Phosphopeptides Using the Q TRAP LC/MS/MS System Purpose This application note describes an automated workflow
More informationSupporting Information
Supporting Information Wiley-VCH 2006 69451 Weinheim, Germany Tissue Imaging at Atmospheric Pressure using Desorption Electrospray Ionization (DESI) Mass Spectrometry Justin M. Wiseman, Demian R. Ifa,
More informationChallenges in Separation Technologies
Challenges in Separation Technologies Erin Shammel Baker Xing Zhang, Yehia Ibrahim, Matthew Monroe, Dennis Mehinagic, Justin Teeguarden, Thomas Metz and Richard D. Smith Pacific Northwest National Laboratory
More informationMass Spectrometry Course Árpád Somogyi Chemistry and Biochemistry MassSpectrometry Facility) University of Debrecen, April 12-23, 2010
Mass Spectrometry Course Árpád Somogyi Chemistry and Biochemistry MassSpectrometry Facility) University of Debrecen, April 12-23, 2010 Introduction, Ionization Methods Mass Analyzers, Ion Activation Methods
More informationCOMBINING LASER CAPTURE MICRODISSECTION AND MALDI MASS SPECTROMETRY FOR TISSUE PROTEIN PROFILING: METHODOLOGY DEVELOPMENT AND CLINICAL APPLICATIONS
COMBINING LASER CAPTURE MICRODISSECTION AND MALDI MASS SPECTROMETRY FOR TISSUE PROTEIN PROFILING: METHODOLOGY DEVELOPMENT AND CLINICAL APPLICATIONS By Baogang Jonathan Xu Dissertation Submitted to the
More informationLOCALISATION, IDENTIFICATION AND SEPARATION OF MOLECULES. Gilles Frache Materials Characterization Day October 14 th 2016
LOCALISATION, IDENTIFICATION AND SEPARATION OF MOLECULES Gilles Frache Materials Characterization Day October 14 th 2016 1 MOLECULAR ANALYSES Which focus? LOCALIZATION of molecules by Mass Spectrometry
More informationFPO. Label-Free Molecular Imaging. Innovation with Integrity. Discover, localize and quantify biochemical changes and molecular markers
FPO Label-Free Molecular Imaging Discover, localize and quantify biochemical changes and molecular markers Innovation with Integrity Mass Spectrometry Bruker, the global leader in MALDI-MS technology,
More informationSolving practical problems. Maria Kuhtinskaja
Solving practical problems Maria Kuhtinskaja What does a mass spectrometer do? It measures mass better than any other technique. It can give information about chemical structures. What are mass measurements
More informationANALYTISCHE STRATEGIE Tissue Imaging. Bernd Bodenmiller Institute of Molecular Life Sciences University of Zurich
ANALYTISCHE STRATEGIE Tissue Imaging Bernd Bodenmiller Institute of Molecular Life Sciences University of Zurich Quantitative Breast single cancer cell analysis Switzerland Brain Breast Lung Colon-rectum
More informationINTRODUCTION TO MALDI IMAGING
INTRODUCTION TO MALDI IMAGING Marten F. Snel, Emmanuelle Claude, Thérèse McKenna, and James I. Langridge Waters Corporation, Manchester, UK INT RODUCTION The last few years have seen a rapid increase in
More informationImaging Mass Microscope
Imaging Mass Microscope imscope C146-E220 Introducing the New Era of Imaging Mass Spectrometry Imaging mass spectrometry is a revolutionary new technology. The instrument is a combination of an optical
More informationApplications for MALDI-TOF Imaging Present and Future
Y Position 8563 / (3000-30000) 12124 / (3000-30000) 14044 / (3000-30000) 10158 35 / (3000-30000) 15012 / (3000-30000) 10894 / (3000-30000) 4964 / (3000-30000) 12414 / (3000-30000) Area Chromatogram - All
More informationThe J105 SIMS. A New Instrument for 3-Dimensional Imaging and Analysis. Paul Blenkinsopp, Ionoptika Ltd
The J105 SIMS A New Instrument for 3-Dimensional Imaging and Analysis Paul Blenkinsopp, Ionoptika Ltd The J105 SIMS Why a new ToF Mass Spectrometer? The J105 ToF has been designed to allow us to separate
More informationApplication Note # MT-91. High Quality MALDI Imaging of Proteins and Peptides in Small Rodent Organ Tissues. Bruker Daltonics.
Bruker Daltonics Application Note # MT-91 18385 Da 6230 Da 7843 Da High Quality MALDI Imaging of Proteins and Peptides in Small Rodent Organ Tissues New developments in MALDI instrumentation, laser technology
More informationLABORATÓRIUMI GYAKORLAT SILLABUSZ SYLLABUS OF A PRACTICAL DEMONSTRATION. financed by the program
TÁMOP-4.1.1.C-13/1/KONV-2014-0001 projekt Az élettudományi-klinikai felsőoktatás gyakorlatorientált és hallgatóbarát korszerűsítése a vidéki képzőhelyek nemzetközi versenyképességének erősítésére program
More informationMALDI imaging mass spectrometry: molecular snapshots of biochemical systems
MALDI imaging mass spectrometry: molecular snapshots of biochemical systems Dale S Cornett, Michelle L Reyzer, Pierre Chaurand & Richard M Caprioli Matrix-assisted laser desorption/ionization (MALDI) imaging
More informationMALDI IMAGING MASS SPECTROMETRY FOR SMALL MOLECULE QUANTITATION AND EVALUATION FOR MARKERS OF DRUG RESPONSE IN GLIOMAS. Sara L. Frappier.
MALDI IMAGING MASS SPECTROMETRY FOR SMALL MOLECULE QUANTITATION AND EVALUATION FOR MARKERS OF DRUG RESPONSE IN GLIOMAS By Sara L. Frappier Dissertation Submitted to the Faculty of the Graduate School of
More informationMALDI-TOF analysis of whole blood: its usefulness and potential in the assessment of HbA1c levels
MALDI-TOF analysis of whole blood: its usefulness and potential in the assessment of HbA1c levels Jane Y. Yang, David A. Herold Department of Pathology, University of California San Diego, 9500 Gilman
More informationDesorption Electrospray Ionization Coupled with Ultraviolet Photodissociation for Characterization of Phospholipid Isomers in Tissue Sections
Desorption Electrospray Ionization Coupled with Ultraviolet Photodissociation for Characterization of Phospholipid Isomers in Tissue Sections Dustin R. Klein, Clara L. Feider, Kyana Y. Garza, John Q. Lin,
More informationMass Spectrometry. - Introduction - Ion sources & sample introduction - Mass analyzers - Basics of biomolecule MS - Applications
- Introduction - Ion sources & sample introduction - Mass analyzers - Basics of biomolecule MS - Applications Adapted from Mass Spectrometry in Biotechnology Gary Siuzdak,, Academic Press 1996 1 Introduction
More informationQualitative & Quantitative MS imaging technique for BAK distribution in eye. Stauber Jonathan, PhD CSO ImaBiotech
Q Qualitative & Quantitative MS imaging technique for BAK distribution in eye Stauber Jonathan, PhD CSO ImaBiotech EBF congress 2012, Bruxelles Q An innovative company with an innovative technology WHAT
More informationLecture 3. Tandem MS & Protein Sequencing
Lecture 3 Tandem MS & Protein Sequencing Nancy Allbritton, M.D., Ph.D. Department of Physiology & Biophysics 824-9137 (office) nlallbri@uci.edu Office- Rm D349 Medical Science D Bldg. Tandem MS Steps:
More informationData Independent MALDI Imaging HDMS E for Visualization and Identification of Lipids Directly from a Single Tissue Section
Data Independent MALDI Imaging HDMS E for Visualization and Identification of Lipids Directly from a Single Tissue Section Emmanuelle Claude, Mark Towers, and Kieran Neeson Waters Corporation, Manchester,
More informationCS 6824: Tissue-Based Map of the Human Proteome
CS 6824: Tissue-Based Map of the Human Proteome T. M. Murali November 17, 2016 Human Protein Atlas Measure protein and gene expression using tissue microarrays and deep sequencing, respectively. Alternative
More informationAutomated Sample Preparation/Concentration of Biological Samples Prior to Analysis via MALDI-TOF Mass Spectroscopy Application Note 222
Automated Sample Preparation/Concentration of Biological Samples Prior to Analysis via MALDI-TOF Mass Spectroscopy Application Note 222 Joan Stevens, Ph.D.; Luke Roenneburg; Tim Hegeman; Kevin Fawcett
More information(III) MALDI instrumentation
Dr. Sanjeeva Srivastava (I) Basics of MALDI-TF (II) Sample preparation In-gel digestion Zip-tip sample clean-up Matrix and sample plating (III) MALDI instrumentation 2 1 (I) Basics of MALDI-TF Analyte
More informationMS/MS to Targeted Proteomics (MRM)
MS/MS to Targeted Proteomics (MRM) How it worked on the Human Lens Proteome Jayson Falkner PhD jay@singleorganism.com Genes Show Limited Value in Predicting Diseases With only a few exceptions, what the
More informationMethods in Mass Spectrometry. Dr. Noam Tal Laboratory of Mass Spectrometry School of Chemistry, Tel Aviv University
Methods in Mass Spectrometry Dr. Noam Tal Laboratory of Mass Spectrometry School of Chemistry, Tel Aviv University Sample Engineering Chemistry Biology Life Science Medicine Industry IDF / Police Sample
More informationShotgun Proteomics MS/MS. Protein Mixture. proteolysis. Peptide Mixture. Time. Abundance. Abundance. m/z. Abundance. m/z 2. Abundance.
Abundance Abundance Abundance Abundance Abundance Shotgun Proteomics Protein Mixture 1 2 3 MS/MS proteolysis m/z 2 3 Time µlc m/z MS 1 m/z Peptide Mixture m/z Block Diagram of a Mass Spectrometer Sample
More informationPUBLISHED VERSION Gustafsson JOR, et al. PERMISSIONS.
PUBLISHED VERSION Gustafsson, Ove Johan Ragnar; McColl, Shaun Reuss; Hoffmann, Peter Imaging mass spectrometry and its methodological application to murine tissue, Journal of Proteomics and Bioinformatics,
More informationIon Source. Mass Analyzer. Detector. intensity. mass/charge
Proteomics Informatics Overview of spectrometry (Week 2) Ion Source Analyzer Detector Peptide Fragmentation Ion Source Analyzer 1 Fragmentation Analyzer 2 Detector b y Liquid Chromatography (LC)-MS/MS
More informationApplication Note # MT-111 Concise Interpretation of MALDI Imaging Data by Probabilistic Latent Semantic Analysis (plsa)
Application Note # MT-111 Concise Interpretation of MALDI Imaging Data by Probabilistic Latent Semantic Analysis (plsa) MALDI imaging datasets can be very information-rich, containing hundreds of mass
More information1. Sample Introduction to MS Systems:
MS Overview: 9.10.08 1. Sample Introduction to MS Systems:...2 1.1. Chromatography Interfaces:...3 1.2. Electron impact: Used mainly in Protein MS hard ionization source...4 1.3. Electrospray Ioniztion:
More informationBiological Mass Spectrometry. April 30, 2014
Biological Mass Spectrometry April 30, 2014 Mass Spectrometry Has become the method of choice for precise protein and nucleic acid mass determination in a very wide mass range peptide and nucleotide sequencing
More information[application note] DIRECT TISSUE IMAGING AND CHARACTERIZATION OF PHOSPHOLIPIDS USING A MALDI SYNAPT HDMS SYSTEM
DIRECT TISSUE IMAGING AND CHARACTERIZATION OF PHOSPHOLIPIDS USING A MALDI SYNAPT HDMS SYSTEM Emmanuelle Claude, Marten Snel, Thérèse McKenna, and James Langridge INTRODUCTION The last decade has seen a
More informationIonization Methods. Neutral species Charged species. Removal/addition of electron(s) Removal/addition of proton(s)
Ionization Methods Neutral species Charged species Removal/addition of electron(s) M + e - (M +. )* + 2e - electron ionization Removal/addition of proton(s) M + (Matrix)-H MH + + (Matrix) - chemical ionization
More informationMass Spectrometry Infrastructure
Mass Spectrometry Infrastructure Todd Williams, Ph.D. Director KU Mass Spectrometry and Analytical Proteomics Laboratory Mass Spectrometry Lab B025 Malott Hall Mission The Mass Spectrometry and analytical
More informationDouble charge of 33kD peak A1 A2 B1 B2 M2+ M/z. ABRF Proteomics Research Group - Qualitative Proteomics Study Identifier Number 14146
Abstract The 2008 ABRF Proteomics Research Group Study offers participants the chance to participate in an anonymous study to identify qualitative differences between two protein preparations. We used
More informationNew Developments in LC-IMS-MS Proteomic Measurements and Informatic Analyses
New Developments in LC-IMS-MS Proteomic Measurements and Informatic Analyses Erin Shammel Baker Kristin E. Burnum-Johnson, Xing Zhang, Cameron P. Casey, Yehia M. Ibrahim, Matthew E. Monroe, Tao Liu, Brendan
More informationIntroduction to Proteomics 1.0
Introduction to Proteomics 1.0 CMSP Workshop Pratik Jagtap Managing Director, CMSP Objectives Why are we here? For participants: Learn basics of MS-based proteomics Learn what s necessary for success using
More informationImprove Protein Analysis with the New, Mass Spectrometry- Compatible ProteasMAX Surfactant
Improve Protein Analysis with the New, Mass Spectrometry- Compatible Surfactant ABSTRACT Incomplete solubilization and digestion and poor peptide recovery are frequent limitations in protein sample preparation
More informationFundamentals of Soft Ionization and MS Instrumentation
Fundamentals of Soft Ionization and MS Instrumentation Ana Varela Coelho varela@itqb.unl.pt Mass Spectrometry Lab Analytical Services Unit Index Mass spectrometers and its components Ionization methods:
More informationSupporting Information MALDI versus ESI the impact of the ion source on peptide identification
Supporting Information MALDI versus ESI the impact of the ion source on peptide identification Wiebke Maria Nadler, 1 Dietmar Waidelich, 2 Alexander Kerner, 1 Sabrina Hanke, 1 Regina Berg, 3 Andreas Trumpp
More informationPhospholipid characterization by a TQ-MS data based identification scheme
P-CN1716E Phospholipid characterization by a TQ-MS data based identification scheme ASMS 2017 MP-406 Tsuyoshi Nakanishi 1, Masaki Yamada 1, Ningombam Sanjib Meitei 2, 3 1 Shimadzu Corporation, Kyoto, Japan,
More information2. Ionization Sources 3. Mass Analyzers 4. Tandem Mass Spectrometry
Dr. Sanjeeva Srivastava 1. Fundamental of Mass Spectrometry Role of MS and basic concepts 2. Ionization Sources 3. Mass Analyzers 4. Tandem Mass Spectrometry 2 1 MS basic concepts Mass spectrometry - technique
More informationA263 A352 A204. Pan CK. pstat STAT3 pstat3 STAT3 pstat3. Columns Columns 1-6 Positive control. Omentum. Rectosigmoid A195.
pstat3 75 Pan CK A A263 A352 A24 B Columns 1-6 Positive control A195 A22 A24 A183 Rectal Nodule STAT3 pstat3 STAT3 pstat3 Columns 7-12 Omentum Rectosigmoid Left Ovary Right Ovary Omentum Uterus Uterus
More informationComparison of mass spectrometers performances
Comparison of mass spectrometers performances Instrument Mass Mass Sensitivity resolution accuracy Quadrupole 1 x 10 3 0.1 Da* 0.5-1.0 pmol DE-MALDI 2 x 10 4 20 ppm 1-10 fmol peptide 1-5 pmol protein Ion
More informationHiroya Hidaka *1), Masaki Takiwaki 2), Mine Yamashita 2), Shinya Otsuki 1), Kenji Kawasaki 3), Mitsutoshi Sugano 3) and Takayuki Honda 4)
Mild acid hydrolysis of sphingolipids yields lysosphingolipids: a matrix-assisted laser desorption and ionization time-of-flight mass spectrometry study Hiroya Hidaka *1), Masaki Takiwaki 2), Mine Yamashita
More informationSupplementary Information
Supplementary Information Molecular imaging of brain localization of liposomes in mice using MALDI mass spectrometry Annabelle Fülöp 1,2, Denis A. Sammour 1,2, Katrin Erich 1,2, Johanna von Gerichten 4,
More informationMass spectrometry has become among the most important
GASTROENTEROLOGY 2012;143:544 549 IMAGING ADVANCED TECHNOLOGY Ralf Kiesslich and Thomas D. Wang, Section Editors Direct Molecular Tissue Analysis by MALDI Imaging Mass Spectrometry in the Field of Gastrointestinal
More informationMetabolite identification in metabolomics: Database and interpretation of MSMS spectra
Metabolite identification in metabolomics: Database and interpretation of MSMS spectra Jeevan K. Prasain, PhD Department of Pharmacology and Toxicology, UAB jprasain@uab.edu utline Introduction Putative
More information9/11/2017. Disclosures. Imaging Mass Spectrometry. Technical Advances and Clinical Problem Solving. High Mass Resolution Imaging (FTICR MS)
Disclosures Academic Affiliation Vanderbilt University School of Medicine MALDI Imaging Mass Spectrometry: Technical Advances and Clinical Problem Solving Industry Affiliations Co-founder & CEO Frontier
More informationCover Page. The handle holds various files of this Leiden University dissertation
Cover Page The handle http://hdl.handle.net/1887/48824 holds various files of this Leiden University dissertation Author: Lou, Sha Title: Biomarker discovery in high grade sarcomas by mass spectrometry
More informationAdvances in Hybrid Mass Spectrometry
The world leader in serving science Advances in Hybrid Mass Spectrometry ESAC 2008 Claire Dauly Field Marketing Specialist, Proteomics New hybrids instruments LTQ Orbitrap XL with ETD MALDI LTQ Orbitrap
More informationNAS Workshop on the Industrialization of Biology
High throughput chemical characterization using mass spectrometry NAS Workshop on the Industrialization of Biology Jonathan V. Sweedler University of Illinois Urbana, IL USA May 2014 Funding: DOE, NSF
More informationNew Instruments and Services
New Instruments and Services Liwen Zhang Mass Spectrometry and Proteomics Facility The Ohio State University Summer Workshop 2016 Thermo Orbitrap Fusion http://planetorbitrap.com/orbitrap fusion Thermo
More informationMatrix Assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry
Matrix Assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry Time-of-Flight Mass Spectrometry. Basic principles An attractive feature of the time-of-flight (TOF) mass spectrometer is its
More informationSUPPORTING INFORMATION. Multimodal Mass Spectrometry Imaging of N-glycans and Proteins from the Same
SUPPORTING INFORMATION Multimodal Mass Spectrometry Imaging of N-glycans and Proteins from the Same Tissue Section. Bram Heijs 1, Stephanie Holst 1, Inge H. Briaire-de Bruijn 2, Gabi W. van Pelt 3, Arnoud
More informationSCS Mass Spectrometry Laboratory
SCS Mass Spectrometry Laboratory Contact Information Staff 31 Noyes Laboratory (8:00-5:00 M-F) 217-333-2545 http://scs.illinois.edu/massspec/ Furong Sun (frs@illinois.edu) Furong Sun Director Training
More informationNext-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry
Nat Met, April 2014 Nat Med, April 2014 Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry Journal Club Timo Böge Overview Introduction Conventional Immunohistochemistry
More informationNormalization in MALDI-TOF imaging datasets of proteins: practical considerations
Anal Bioanal Chem (2011) 401:167 181 DOI 10.1007/s00216-011-4929-z ORIGINAL PAPER Normalization in MALDI-TOF imaging datasets of proteins: practical considerations Sören-Oliver Deininger & Dale S. Cornett
More informationBiological Mass spectrometry in Protein Chemistry
Biological Mass spectrometry in Protein Chemistry Tuula Nyman Institute of Biotechnology tuula.nyman@helsinki.fi MASS SPECTROMETRY is an analytical technique that identifies the chemical composition of
More informationProtein Analysis using Electrospray Ionization Mass Spectroscopy *
OpenStax-CNX module: m38341 1 Protein Analysis using Electrospray Ionization Mass Spectroscopy * Wilhelm Kienast Andrew R. Barron This work is produced by OpenStax-CNX and licensed under the Creative Commons
More informationUsing Infrared and Raman Microspectroscopies to Compare Ex Vivo Involved Psoriatic Skin and Normal Human Skin
Using Infrared and Raman Microspectroscopies to Compare Ex Vivo Involved Psoriatic Skin and Normal Human Skin Leroy, Marie (Laboratoire d'ingénierie de Surface (LIS)) Lefèvre, Thierry (Groupe de recherche
More informationO O H. Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION EXPERIMENTAL. LC /MS conditions
Simplifying Qual/Quan Analysis in Discovery DMPK using UPLC and Xevo TQ MS Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION The determination of the drug metabolism
More informationDetection, Differentiation, and Subtyping of Botulinum Neurotoxins with Mass Spectrometry
Detection, Differentiation, and Subtyping of Botulinum Neurotoxins with Mass Spectrometry Suzanne R. Kalb*, Hercules Moura*, Wanda I. Santana*, Jakub Baudys*, Maria I. Solano*, Adrian R. Woolfitt*, Theresa
More informationMT09 - Normal Human Tissue Microarray, FDA
Reveal Biosciences offers Histochemical Staining, Immunohistochemistry (IHC), In Situ Hybridization (ISH), Whole Slide Imaging, and Quantitative Image Analysis on any TMA MT09 - Normal Human Tissue Microarray,
More informationPosterREPRINT AN AUTOMATED METHOD TO SELF-CALIBRATE AND REJECT NOISE FROM MALDI PEPTIDE MASS FINGERPRINT SPECTRA
Overview AN AUTOMATED METHOD TO SELF-CALIBRATE AND REJECT NOISE FROM MALDI PEPTIDE MASS FINGERPRINT SPECTRA Jeffery M Brown, Neil Swainston, Dominic O. Gostick, Keith Richardson, Richard Denny, Steven
More informationMass Spectrometry. Actual Instrumentation
Mass Spectrometry Actual Instrumentation August 2017 See also http://www.uni-bielefeld.de/chemie/analytik/ms f additional infmation 1. MALDI TOF MASS SPECTROMETRY ON THE ULTRAFLEX 2 2. ESI MASS SPECTROMETRY
More informationStructural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry
PO-CON1347E Structural Elucidation of N-glycans Originating From Ovarian Cancer Cells Using High-Vacuum MALDI Mass Spectrometry ASMS 2013 TP-708 Matthew S. F. Choo 1,3 ; Roberto Castangia 2 ; Matthew E.
More informationAnalysis of Triglycerides in Cooking Oils Using MALDI-TOF Mass Spectrometry and Principal Component Analysis
Analysis of Triglycerides in Cooking Oils Using MALDI-TOF Mass Spectrometry and Principal Component Analysis Kevin Cooley Chemistry Supervisor: Kingsley Donkor 1. Abstract Triglycerides are composed of
More informationREDOX PROTEOMICS. Roman Zubarev.
REDOX PROTEOMICS Roman Zubarev Roman.Zubarev@ki.se Physiological Chemistry I, Department for Medical Biochemistry & Biophysics, Karolinska Institutet, Stockholm What is (RedOx) Proteomics? Proteomics -
More informationHigh Resolution Glycopeptide Mapping of EPO Using an Agilent AdvanceBio Peptide Mapping Column
High Resolution Glycopeptide Mapping of EPO Using an Agilent AdvanceBio Peptide Mapping Column Application Note BioPharma Authors James Martosella, Phu Duong, and Alex Zhu Agilent Technologies, Inc. Abstract
More informationFOCUS: MALDI-TOF AND BIOLOGICAL MS: RESEARCH ARTICLE
B American Society for Mass Spectrometry, 2011 J. Am. Soc. Mass Spectrom. (2011) 22:1022Y1031 DOI: 10.1007/s13361-011-0121-0 FOCUS: MALDI-TOF AND BIOLOGICAL MS: RESEARCH ARTICLE High-Speed MALDI-TOF Imaging
More informationNew Instruments and Services
New Instruments and Services http://planetorbitrap.com/orbitrap fusion Combining the best of quadrupole, Orbitrap, and ion trap mass analysis in a revolutionary Tribrid architecture, the Orbitrap Fusion
More informationHPLC-MALDI-TOF Mass Spectrometry for the Analysis of Complex Polymers
TU Darmstadt HPLC-MALDI-TOF Mass Spectrometry for the Analysis of Complex Polymers H. Pasch German Institute for Polymers (Deutsches Kunststoff-Institut) Darmstadt, Germany hpasch@dki.tu-darmstadt.de 2004
More informationMass Spectrometry based metabolomics
Mass Spectrometry based metabolomics Metabolomics- A realm of small molecules (
More information