imsresearch.med.helsinki.fi/corefacilities/proteinchem... · 2012-10-08 · benefits of maldi-ims...
TRANSCRIPT
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IMSImaging mass spectrometry
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A 384 position MALDI-TOF sample target plate. To each position 0.5-2µl of sample together with matrix solution is pipetted and allowed to dry.
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PrinciplesTissue section (mouse brain)
2,000 15,000 30,000
Ion
inte
nsity
Acquisition x
Acqu
isitio
n y
m/z
12mm
• A laser is rastered over a defined area while acquiring a complete mass spectrum from each position, resulting in molecular images for multiple analytesCornett, et al., Nature Methods 2007
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IMS workflow
Stoeckli, M, et al., Analytical Biochemistry 2002
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Tissue slide for IMS
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Bruker IMAGE-PREPWith SPR
Nano spotter
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Benefits of MALDI-IMS
• Analysis of entire sample in one reading
• Previous knowledge of molecular composition is not necessary
• Allows for investigation of disease formation, progression, and treatment
www.maldi-msi.org
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A whole mouse
Penetration of the drug into the tissue
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MS imaging advantages
•No labeling requiredBiomolecules are functionally unmodified
•Image biomolecular modificationsPTM’s, Metabolites
•Detailed information on molecular identityBy 2D MALDI
•Large scope of different elements and molecules
Targeted labeling
Label free imaging
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IMS vs. histochemical stain
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New Phytologist (2007) 173 : 438 –444m/z 381
m/z 543
m/z 705
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Reconstruction of the Carbohydrate moietyOf WHEAT (Triticum aestivum)
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3D IMS-MS
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Alois Alzheimer’s disease
Alois Alzheimer
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Alzheimer’s Amyloid Precursor Protein (APP)
sA
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?
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Microchips on IMS
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Chemical sensor arrays for application in mass spectrometry on surfaces and
tissues
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Multiplex Targeted Secondary Detection(MTSD)
• Targeting molecule of interest with specific antibodies which carry a photocleavable mass tag sensor
• The mass tag sensor is a small molecule of known mass easily detectable by MALDI MS (indirect detection)
• The tag is released just before detection step due to irradiation with MALDI laser (photolabile linker)
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Thiery et al. Proteomics 2008, 8, 3725–3734
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„normal“ MTSD Arrays
• Samples will be spotted on chip surface and immobilized
• Chip will be treated with mass tagged antibodies
• Matrix application• Under MS conditions: sensor tags will be
released and detected via TOF
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Inverse MTSD Arrays• Antibodies are attached to chip surface• Sample is applied on chip, specific substrates
bind to their antibodies• Matrix application• Substrate-antibody complex gets released and
detected via TOF h.h.h.
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Resolving power of an MS
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IMS FT-ICR-MS
Sensitivity in the zmol range
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Peptide digestion on the membrane (identification of the proteins)
Direct trypsin spraying; (Jardin-Mathé et al., 2008)
2D- IMS for protein identification
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Peptide digestion on the membrane (identification
of the proteins)
Other use of...
- Laser Capture MC of single cells- protein profiling directly from gelsetc
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Imaging Mass Spectrometry (IMS) of a Specific Fragment of Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Kinase Kinase 2 Discriminates Cancer from Uninvolved Prostate Tissue
Lisa H. Cazares,1,2,3 Dean Troyer,1,3 Savvas Mendrinos,3 Raymond A. Lance,3,5, Julius O. Nyalwidhe,1,2,3 Hind A. Beydoun,4 Mary Ann Clements,1,2,3, Richard R. Drake,1,2,3 and O. John Semmes1,2,3
Clin Cancer Res 2009;15(17) September 1, 2009
Diagnostic value?
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MALDI mass spectrometric imaging of biological tissue sections
Tatiana C. Rohner, Dieter Staab and Markus StoeckliNovartis Institutes for BioMedical Research, Novartis Pharma AG, Lichtstrasse 35, CH-4002 Basel, Switzerland
Mechanisms of Ageing and Development Volume 126, Issue 1 , January 2005, Pages 177-185
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What else can we do?
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SELDI = Chip based surface enchanced laser desorption analysis
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The Chip
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Current Developments In SELDI Affinity TechnologyMass Spectrometry Reviews, 23, 34-44, (2004)
High Throughput
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Unraveling biological pathways using the InteractionDiscovery Mapping™ platform
Whole Cell analysis
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Whole Cell and Protein Microarray
Chip structures
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Cell trapping and - lysis Chip structures
Development of Integrated Nanoliter Analysis Devices (DDTC-Viikki, Microtechnology Center-HUT, Biomedicum Helsinki)
Cell trapping chamber
Solvent flushing to MS
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Whole Cell or single Cell
Proteomics!
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Key word: microfluidistics?
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Microfluidics in a Chip for Proteomics
Biochip-IMB, Ltd.
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Biochip-IMB, Ltd.
Express diagnostics tools CUSTOMIZED
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Direct on-chip mass-spectrometry
Biotinylated proteins are bound to avidin immobilized in gel pad.
Protein-ligand interactions
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A typical Chip and how toMake it...
Experion Automated Electrophoresis Station
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The MOTC-nESI Lab-on-a-Chip
Evangelos Gogolides, Athens
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NanoSpray LC/MS
TriVersa™ NanoMate® and ESI Chip® System
Advion Biosiences
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NanoSpray LC/MS
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New nebulizers for Proteomic analyses
Equipped with active surfaces
LC chip test assembly; bead packed silicon column in optical microscope
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2D-CE/MS/MS
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High throughput CE
10000 Capillaries
Capillary windows
Autosampler
LED detection (MIT, Forest et al, 2006)
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Microarray MS Chip (IonTrap)
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A set of thousands of MassSpectrometers in one chip