FPO. Label-Free Molecular Imaging. Innovation with Integrity. Discover, localize and quantify biochemical changes and molecular markers

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1 FPO Label-Free Molecular Imaging Discover, localize and quantify biochemical changes and molecular markers Innovation with Integrity Mass Spectrometry

2 Bruker, the global leader in MALDI-MS technology, is also your partner for label-free molecular MALDI Imaging. MALDI is the most robust ion desorption technology for analyzing many different classes of analyte molecules from many types of samples. When MALDI spectra are collected in a spatially orientated pattern, each spectrum represents a molecular fingerprint of a specific and unique region of the sample being analyzed. Any detected ion can be projected into a 2 dimensional map of sample location and intensity an ion image. From a single dataset, hundreds to thousands of unique, label-free ion images may be generated and used for mapping targeted compounds or untargeted discovery of molecular markers which have a specific regional localization. MALDI Imaging Ion images of six uniquely distributed phospholipids from the below rat testis section Biomarker discovery or targeted monitoring MALDI Imaging is a powerful label-free molecular imaging tool for biomarker discovery or targeted monitoring. This revolutionary technique reveals molecular distributions on a cellular scale and can be a key problem-solver in many different fields of research: Lipidomics Proteomics Drug discovery Cellular disease Infectious disease Plant metabolism Entomology Inorganic surfaces Benefits: Add a spatial component to LC-based omics studies Discover regionally specific biochemical changes and molecular markers Localize and quantify for drug discovery Add molecular specificity to other imaging techniques Correlate protein glycosylations to disease Visually explore metabolic pathways *Cover image courtesy of Dr. Jeff Spraggins, Mass Spectrometry Research Center at Vanderbilt University

3 Add a spatial component to LC-based omics Homogenized samples analyzed by LC-MS/MS present no localization information and the process may dilute highly localized compounds below limit of detection. Analyze samples by MALDI Imaging and capture compound distributions, even when localized to very small compartments. Proteins Lipids Metabolites... MALDI Imaging can simultaneously map hundreds of different compounds within a small region or across large cohorts of samples. Locate, Discover, Identify.. Analyze many samples by MALDI Imaging to identify regionally-specific molecular changes. Match these molecules against LC-MS/MS data from the same samples for confirmation and identification. Analyze many samples by LC-MS/MS to identify sample-specific molecular changes. Map these molecules in the samples using MALDI Imaging to confirm that they have proper regional specificity. Locate 3 potential biomarkers and correlate with pathology Discover 3 potential sample-specific biomarker ions Homogenize Sample #2 Compare LC-MS/MS analysis Sample #1

4 Correlate protein glycosylation to disease N-Glycan maps show glycolylations specific to colorectal cancer PNGase F treatment liberates N-linked glycans Visualize patterns of glycosylation Identify N-glycans using MS/MS Correlate glycosylation patters to pathology Tryptic peptides map protein localization in colorectal cancer Trypsin cleaves intractable proteins into detectable peptides Tryptic peptides reveal distribution of larger proteins in fresh frozen and FFPE tissues Identify protein from peptide MS/MS Access insoluble proteins Richard R. Drake, Professor & Director of MUSC Proteomics Center We have developed and continue to evolve new glycan and glycoprotein methods for MALDI imaging. Bruker has been a strong partner in facilitating and supporting these efforts with their innovative instrumentation and software solutions.

5 Discover regionally specific biochemical differences and molecular markers Cellular heterogeneity of lymphoma is reflected in the SCiLS Lab segmentation map where cellular regions which express similar molecular profiles are presented in the same color. PCA analysis 0.88 AUC ROC analysis Dendrogram and segmentation map shows regions of higher molecular similarity in same color. The segmentation map correlates very well with tissue pathology and allows one to analyze those regions of highest interest with greater specificity. * Sample courtesy of Prof. Etienne Waelkens, KU Leuven, Belgium

6 Systems for highest sample throughput Flex MALDI-TOF systems offer the most versatile analytical range and speed while producing hundreds of ion images per sample. autoflex max: outstanding entry-level MALDI system for Imaging and much more. TOF/TOF is available. Layer of purkinje cells visible ultraflextreme: High performance TOF/TOF imaging system 3 phospholipids mapped from rat brain section ultraflextreme: High performance TOF/TOF imaging system rapiflex: novel laser optics designed specifically for highest performance and highest speed for MALDI Imaging. TOF/TOF is available.

7 Systems for highest information content scimax and 2XR MRMS systems offer the highest mass resolving power of any commercial MALDI system and are the ultimate systems for imaging drugs, metabolites and lipids, producing thousands of unique images per sample. Realize the full benefit of extreme Resolution. Above, a 3-color image from rat testis showing distributions of three ions at nominal m/z 848 and differing only by 300 mda. Below, for smaller molecules, extreme Resolution can reveal high-fidelity Isotopic Fine Structure that can be used to assign unique molecular formula identification. SmartFormula hits for measured m/z ± 1ppm A2 Peak, measured * 34 S A2 peak, theoretical for C 14 H 20 N 4 S Isotopic Fine Structure of A2 matches the theoretical IFS pattern of the correct formula 13 C 2 13 C 15 N 13 C 33 S m/z *Interfering ion

8 Localize and quantify for drug discovery Make critical decisions earlier: The low operational cost of MALDI Imaging allows it to be deployed very early in the discovery pipeline to help make decisions faster and bridges the gap between traditional LC-MS and Quantitative Whole Body Autoradiography (QWBA) for distribution and metabolism studies. Whether part of dosing and PK studies or for ADME/Tox studies, MALDI Imaging offers high sensitivity, multiple target localization, and simultaneous discovery of toxicity markers all integrated with histology. M M-CH 3 MOH % Gd of injected dose Benefit/Technique QWBA MALDI Imaging LC-MS/MS Spatial distribution X Drug/metabolite differentiation X Quantitative Integrate with histology X Label free X Steve Castellino, Head of US ex vivo imaging and Senior GSK Fellow We have been able to achieve the spatial and spectral resolution required to examine sub-compartment tissue distributions and correlate them with histology in the preclinical setting. This ability to link chemistry and biology is permitting us to more closely examine the basis of drug toxicity and pharmacology as well as refine our understanding of pharmacokinetics and drug transport.

9 Visually explore biochemical pathways Control Parkinson's L D M R NH 2 Room temperature, ph>7 N H 2O R V 10 mm DA 3-MT 5-HT GABA Dopamine (DA) GABA glutamate * Images courtesy of Prof. Per Andrén, Uppsala Univ. Prof. Per Andrén, Uppsala University, Sweden Mass spectrometry imaging enables us to simultaneously map and quantitate multiple neurotransmitters, their precursors and metabolites directly in tissue sections. That is, almost the complete dopaminergic and serotonergic neurotransmitter networks.

10 Complete software toolkit Acquire Visualize Analyze Information Acquire: Method driven image acquisition Define measurement areas from microscopic image Image all of the sample or just regions of interest Visualize: Multiple ion overlays ROI segmentation Save targeted ions for instant visualization or explore interactively Analyze: SCiLS Lab Core: auto-segmentation and co-localization SCiLS Cloud integrates remote teams SCiLS Lab Pro: full analyses of large sample cohorts SCiLS Lab Premium 3D: full analyses of 2- and 3-dimensional images

11 Add molecular specificity to other imaging techniques Fluorescence Look beyond optical morphology. Import many other imaging modalities to examine your MSI data in the most relevant context and boost confidence in your analyses. Histology Histology* Whole Tissue MALDI Imaging* Atlas Reference IHC Tissue microarrays (TMA) Bruker pioneered the integration of MALDI Imaging data with other imaging modalities to provide you with the most biologically relevant information. Bruker s imaging software allows co-registration and overlays of multiple images with full user control. * Image courtesy of Dr. Jeff Spraggins, Mass Spectrometry Research Center at Vanderbilt, USA

12 Bruker Daltonics is continually improving its products and reserves the right to change specifications without notice. BDAL , Contact your local Bruker representative to learn more about our MALDI imaging products and how they can help you gain a deeper understanding of your samples. scimax & 2XR rapiflex ultraflextreme autoflex max For the highest spatial resolution Bruker offers the industry leading TM Sprayer from HTX Imaging Dr. Jonathan Stauber, CEO Imabiotech ImaBiotech purchased several Bruker instruments because their devices high performance fit perfectly with our MALDI Imaging technology platform to provide superior services to the pharmaceutical and biomedical industries. Our extensive research made possible by integrating Bruker into our repertoire is proof of the unparalleled quality and robustness of Bruker technology. For research use only. Not for use in diagnostic procedures. Bruker Daltonik GmbH Bremen Germany Phone 49 (0) Fax 49 (0) Bruker Daltonics Inc. Billerica, MA USA Phone 1 (978) Fax 1 (978) ms.sales.bdal@bruker.com - Scan the QR-Code for more Details