IMSImaging mass spectrometry

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.

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

IMS workflow

Stoeckli, M, et al., Analytical Biochemistry 2002

Tissue slide for IMS

Bruker IMAGE-PREPWith SPR

Nano spotter

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

A whole mouse

Penetration of the drug into the tissue

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

IMS vs. histochemical stain

New Phytologist (2007) 173 : 438 –444m/z 381

m/z 543

m/z 705

Reconstruction of the Carbohydrate moietyOf WHEAT (Triticum aestivum)

3D IMS-MS

Alois Alzheimer’s disease

Alois Alzheimer

Alzheimer’s Amyloid Precursor Protein (APP)

sA

?

Microchips on IMS

Chemical sensor arrays for application in mass spectrometry on surfaces and

tissues

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)

Thiery et al. Proteomics 2008, 8, 3725–3734

„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

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.

Resolving power of an MS

IMS FT-ICR-MS

Sensitivity in the zmol range

Peptide digestion on the membrane (identification of the proteins)

Direct trypsin spraying; (Jardin-Mathé et al., 2008)

2D- IMS for protein identification

Peptide digestion on the membrane (identification

of the proteins)

Other use of...

- Laser Capture MC of single cells- protein profiling directly from gelsetc

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?

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

What else can we do?

SELDI = Chip based surface enchanced laser desorption analysis

The Chip

Current Developments In SELDI Affinity TechnologyMass Spectrometry Reviews, 23, 34-44, (2004)

High Throughput

Unraveling biological pathways using the InteractionDiscovery Mapping™ platform

Whole Cell analysis

Whole Cell and Protein Microarray

Chip structures

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

Whole Cell or single Cell

Proteomics!

Key word: microfluidistics?

Microfluidics in a Chip for Proteomics

Biochip-IMB, Ltd.

Biochip-IMB, Ltd.

Express diagnostics tools CUSTOMIZED

Direct on-chip mass-spectrometry

Biotinylated proteins are bound to avidin immobilized in gel pad.

Protein-ligand interactions

A typical Chip and how toMake it...

Experion Automated Electrophoresis Station

The MOTC-nESI Lab-on-a-Chip

Evangelos Gogolides, Athens

NanoSpray LC/MS

TriVersa™ NanoMate® and ESI Chip® System

Advion Biosiences

NanoSpray LC/MS

New nebulizers for Proteomic analyses

Equipped with active surfaces

LC chip test assembly; bead packed silicon column in optical microscope

2D-CE/MS/MS

High throughput CE

10000 Capillaries

Capillary windows

Autosampler

LED detection (MIT, Forest et al, 2006)

Microarray MS Chip (IonTrap)

A set of thousands of MassSpectrometers in one chip