35 results on '"Richard J A, Goodwin"'
Search Results
2. Method to Investigate the Distribution of Water-Soluble Drug-Delivery Systems in Fresh Frozen Tissues Using Imaging Mass Cytometry
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Richard M. England, Richard J. A. Goodwin, Stephanie Ling, Jonathan A. Rose, Nicole Strittmatter, Marianne Ashford, Ian Purvis, Alan M. Race, Daniel Sutton, Jennifer I. Moss, Edmond Wong, Ruth Macdonald, Simon T. Barry, and Gareth Maglennon
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Staining and Labeling ,Chemistry ,medicine.medical_treatment ,010401 analytical chemistry ,Water ,010402 general chemistry ,Ablation ,01 natural sciences ,Mass Spectrometry ,0104 chemical sciences ,Analytical Chemistry ,Matrix (chemical analysis) ,Drug Delivery Systems ,Water soluble drug ,Dendrimer ,Immunochemistry ,Fresh frozen ,medicine ,Distribution (pharmacology) ,Mass cytometry ,Image Cytometry ,Biomedical engineering - Abstract
Imaging mass cytometry (IMC) offers the opportunity to image metal- and heavy halogen-containing xenobiotics in a highly multiplexed experiment with other immunochemistry-based reagents to distinguish uptake into different tissue structures or cell types. However, in practice, many xenobiotics are not amenable to this analysis, as any compound which is not bound to the tissue matrix will delocalize during aqueous sample-processing steps required for IMC analysis. Here, we present a strategy to perform IMC experiments on a water-soluble polysarcosine-modified dendrimer drug-delivery system (S-Dends). This strategy involves two consecutive imaging acquisitions on the same tissue section using the same instrumental platform, an initial laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MSI) experiment followed by tissue staining and a standard IMC experiment. We demonstrated that settings can be found for the initial ablation step that leave sufficient residual tissue for subsequent antibody staining and visualization. This workflow results in lateral resolution for the S-Dends of 2 μm followed by imaging of metal-tagged antibodies at 1 μm.
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- 2021
3. Mass Spectrometry Detection and Imaging of a Non-Covalent Protein-Drug Complex in Tissue from Orally Dosed Rats
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Eva Illes‐Toth, Oliver J. Hale, James W. Hughes, Nicole Strittmatter, Jonathan Rose, Ben Clayton, Rebecca Sargeant, Stewart Jones, Andreas Dannhorn, Richard J. A. Goodwin, and Helen J. Cooper
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Diagnostic Imaging ,Liver ,Drug Discovery ,Animals ,General Chemistry ,General Medicine ,Bezafibrate ,Catalysis ,Mass Spectrometry ,Rats - Abstract
Here, we demonstrate detection by mass spectrometry of an intact protein-drug complex directly from liver tissue from rats that had been orally dosed with the drug. The protein-drug complex comprised fatty acid binding protein 1, FABP1, non-covalently bound to the small molecule therapeutic bezafibrate. Moreover, we demonstrate spatial mapping of the [FABP1+bezafibrate] complex across a thin section of liver by targeted mass spectrometry imaging. This work is the first demonstration of in situ mass spectrometry analysis of a non-covalent protein-drug complex formed in vivo and has implications for early stage drug discovery by providing a route to target-drug characterization directly from the physiological environment.
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- 2022
4. Revealing the Regional Localization and Differential Lung Retention of Inhaled Compounds by Mass Spectrometry Imaging
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Gregory Hamm, Richard J. A. Goodwin, Nicole Strittmatter, Markus Fridén, Mikael Brülls, Anna Nilsson, Erica Bäckström, Per E. Andrén, and Ken Grime
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Male ,Pulmonary and Respiratory Medicine ,Drug ,media_common.quotation_subject ,Pharmaceutical Science ,Pharmacology ,Mass Spectrometry ,Fluticasone propionate ,Drug Delivery Systems ,Pharmacokinetics ,Administration, Inhalation ,medicine ,Animals ,Distribution (pharmacology) ,Albuterol ,Tissue Distribution ,Pharmacology (medical) ,Rats, Wistar ,Lung ,Salmeterol Xinafoate ,media_common ,Inhalation ,business.industry ,respiratory system ,Bronchodilator Agents ,Rats ,respiratory tract diseases ,medicine.anatomical_structure ,Salbutamol ,Fluticasone ,Salmeterol ,business ,medicine.drug - Abstract
Background: For the treatment of respiratory disease, inhaled drug delivery aims to provide direct access to pharmacological target sites while minimizing systemic exposure. Despite this long-held tenet of inhaled therapeutic advantage, there are limited data of regional drug localization in the lungs after inhalation. The aim of this study was to investigate the distribution and retention of different chemotypes typifying available inhaled drugs [slowly dissolving neutral fluticasone propionate (FP) and soluble bases salmeterol and salbutamol] using mass spectrometry imaging (MSI). Methods: Salmeterol, salbutamol, and FP were simultaneously delivered by inhaled nebulization to rats. In the same animals, salmeterol-d3, salbutamol-d3, and FP-d3 were delivered by intravenous (IV) injection. Samples of lung tissue were obtained at 2- and 30-minute postdosing, and high-resolution MSI was used to study drug distribution and retention. Results: IV delivery resulted in homogeneous lung distribution for all molecules. In comparison, while inhalation also gave rise to drug presence in the entire lung, there were regional chemotype-dependent areas of higher abundance. At the 30-minute time point, inhaled salmeterol and salbutamol were preferentially retained in bronchiolar tissue, whereas FP was retained in all regions of the lungs. Conclusion: This study clearly demonstrates that inhaled small molecule chemotypes are differentially distributed in lung tissue after inhalation, and that high-resolution MSI can be applied to study these retention patterns.
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- 2020
5. Holistic Characterization of a
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Nicole, Strittmatter, Panchali, Kanvatirth, Paolo, Inglese, Alan M, Race, Anna, Nilsson, Andreas, Dannhorn, Hiromi, Kudo, Robert D, Goldin, Stephanie, Ling, Edmond, Wong, Frank, Seeliger, Maria Paola, Serra, Scott, Hoffmann, Gareth, Maglennon, Gregory, Hamm, James, Atkinson, Stewart, Jones, Josephine, Bunch, Per E, Andrén, Zoltan, Takats, Richard J A, Goodwin, and Pietro, Mastroeni
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Mice, Inbred C57BL ,Salmonella typhimurium ,Mice ,Salmonella Infections ,Animals ,Female ,Mass Spectrometry ,Molecular Imaging - Abstract
A more complete and holistic view on host-microbe interactions is needed to understand the physiological and cellular barriers that affect the efficacy of drug treatments and allow the discovery and development of new therapeutics. Here, we developed a multimodal imaging approach combining histopathology with mass spectrometry imaging (MSI) and same section imaging mass cytometry (IMC) to study the effects of
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- 2021
6. Holistic characterization of a salmonella typhimurium infection model using integrated molecular imaging
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Nicole Strittmatter, Gregory Hamm, Stephanie Ling, Scott Hoffmann, Josephine Bunch, Panchali Kanvatirth, Robert D. Goldin, Frank Seeliger, Gareth Maglennon, Richard J. A. Goodwin, Stewart Jones, Alan M. Race, Zoltan Takats, Hiromi Kudo, Andreas Dannhorn, Edmond Wong, Maria Paola Serra, James Atkinson, Anna Nilsson, Pietro Mastroeni, Paolo Inglese, Per E. Andrén, Strittmatter, Nicole [0000-0003-1277-9608], Inglese, Paolo [0000-0001-6179-9643], Race, Alan M [0000-0001-8996-2641], Dannhorn, Andreas [0000-0002-1087-4057], Ling, Stephanie [0000-0002-1237-091X], Andrén, Per E [0000-0002-4062-7743], Takats, Zoltan [0000-0002-0795-3467], and Apollo - University of Cambridge Repository
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Salmonella typhimurium ,Salmonella ,Cell type ,imaging mass cytometry ,mass spectrometry imaging ,medicine.disease_cause ,Stem cell marker ,Mass Spectrometry ,Mass spectrometry imaging ,Salmonella infection ,Analytical Chemistry ,Mice ,Immune system ,Structural Biology ,medicine ,Animals ,multimodal data integration ,Mass cytometry ,network analysis ,Spectroscopy ,0306 Physical Chemistry (incl. Structural) ,0304 Medicinal and Biomolecular Chemistry ,Chemistry ,Phenotype ,Molecular Imaging ,Cell biology ,Mice, Inbred C57BL ,Salmonella Infections ,Female ,Molecular imaging ,0301 Analytical Chemistry - Abstract
A more complete and holistic view on host-microbe interactions is needed to understand the physiological and cellular barriers that affect the efficacy of drug treatments and allow the discovery and development of new therapeutics. Here, we developed a multimodal imaging approach combining histopathology with mass spectrometry imaging (MSI) and same section imaging mass cytometry (IMC) to study the effects of Salmonella Typhimurium infection in the liver of a mouse model using the S. Typhimurium strains SL3261 and SL1344. This approach enables correlation of tissue morphology and specific cell phenotypes with molecular images of tissue metabolism. IMC revealed a marked increase in immune cell markers and localization in immune aggregates in infected tissues. A correlative computational method (network analysis) was deployed to find metabolic features associated with infection and revealed metabolic clusters of acetyl carnitines, as well as phosphatidylcholine and phosphatidylethanolamine plasmalogen species, which could be associated with pro-inflammatory immune cell types. By developing an IMC marker for the detection of Salmonella LPS, we were further able to identify and characterize those cell types which contained S. Typhimurium.
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- 2021
7. Correlative Imaging of Trace Elements and Intact Molecular Species in a Single-Tissue Sample at the 50 μm Scale
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Roger P. Webb, Richard J. A. Goodwin, Josephine Bunch, Alex Dexter, Melanie J. Bailey, Amy Burton, Firat Kaya, Catia Costa, Chelsea J. Nikula, Véronique Dartois, Vladimir Palitsin, Janella de Jesus, Mark A. Chambers, and Adam J. Taylor
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0303 health sciences ,Desorption electrospray ionization ,Spectrometry, Mass, Electrospray Ionization ,Chemistry ,Electrospray ionization ,010401 analytical chemistry ,Analytical chemistry ,Tissue sample ,Mass spectrometry ,01 natural sciences ,Lipids ,Mass spectrometry imaging ,0104 chemical sciences ,Analytical Chemistry ,Molecular Imaging ,Trace Elements ,03 medical and health sciences ,Matrix-assisted laser desorption/ionization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Correlative imaging ,Molecular imaging ,Sulfur ,030304 developmental biology - Abstract
Elemental and molecular imaging play a crucial role in understanding disease pathogenesis. To accurately correlate elemental and molecular markers, it is desirable to perform sequential elemental and molecular imaging on a single-tissue section. However, very little is known about the impact of performing these measurements in sequence. In this work, we highlight some of the challenges and successes associated with performing elemental mapping in sequence with mass spectrometry imaging. Specifically, the feasibility of molecular mapping using the mass spectrometry imaging (MSI) techniques matrix-assisted laser desorption ionization (MALDI) and desorption electrospray ionization (DESI) in sequence with the elemental mapping technique particle-induced X-ray emission (PIXE) is explored. Challenges for integration include substrate compatibility, as well as delocalization and spectral changes. We demonstrate that while sequential imaging comes with some compromises, sequential DESI-PIXE imaging is sufficient to correlate sulfur, iron, and lipid markers in a single tissue section at the 50 μm scale.
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- 2021
8. Comparison of c-13 mri of hyperpolarized [1-c-13]pyruvate and lactate with the corresponding mass spectrometry images in a murine lymphoma model
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Dominique-Laurent Couturier, Vencel Somai, Maria Fala, Simon T. Barry, Kevin M. Brindle, Richard L. Hesketh, Gregory Hamm, Richard J. A. Goodwin, Josephine Bunch, Alan J. Wright, Andreas Dannhorn, Katherine Gibson, Zoltan Takats, INSERM, Université de Lille, University of Cambridge [UK] [CAM], Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM) - U1192, AstraZeneca [Cambridge, UK], Imperial College London, University of Cambridge [UK] (CAM), Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM) - U 1192 (PRISM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)
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tumor ,Lymphoma ,[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging ,pyruvate ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,mass spectrometry imaging ,Mass spectrometry ,Mass Spectrometry ,Mass spectrometry imaging ,030218 nuclear medicine & medical imaging ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Nuclear magnetic resonance ,hyperpolarized 13C MRI ,Murine tumor ,In vivo ,Pyruvic Acid ,Animals ,Radiology, Nuclear Medicine and imaging ,Lactic Acid ,Pyruvate Metabolism ,Note—Preclinical and Clinical Imaging ,Carbon Isotopes ,lactate ,Murine lymphoma ,Chemistry ,Note ,Magnetic Resonance Imaging ,Mass spectrometric ,hyperpolarized C-13 MRI ,030217 neurology & neurosurgery ,Ex vivo - Abstract
International audience; PURPOSE: To compare carbon-13 ((13) C) MRSI of hyperpolarized [1-(13) C]pyruvate metabolism in a murine tumor model with mass spectrometric (MS) imaging of the corresponding tumor sections in order to cross validate these metabolic imaging techniques and to investigate the effects of pyruvate delivery and tumor lactate concentration on lactate labeling.METHODS: [1-(13) C]lactate images were obtained from tumor-bearing mice, following injection of hyperpolarized [1-(13) C]pyruvate, using a single-shot 3D (13) C spectroscopic imaging sequence in vivo and using desorption electrospray ionization MS imaging of the corresponding rapidly frozen tumor sections ex vivo. The images were coregistered, and levels of association were determined by means of Spearman rank correlation and Cohen kappa coefficients as well as linear mixed models. The correlation between [1-(13) C]pyruvate and [1-(13) C]lactate in the MRS images and between [(12) C] and [1-(13) C]lactate in the MS images were determined by means of Pearson correlation coefficients.RESULTS: [1-(13) C]lactate images generated by MS imaging were significantly correlated with the corresponding MRS images. The correlation coefficient between [1-(13) C]lactate and [1-(13) C]pyruvate in the MRS images was higher than between [1-(13) C]lactate and [(12) C]lactate in the MS images.CONCLUSION: The inhomogeneous distribution of labeled lactate observed in the MRS images was confirmed by MS imaging of the corresponding tumor sections. The images acquired using both techniques show that the rate of (13) C label exchange between the injected pyruvate and endogenous tumor lactate pool is more correlated with the rate of pyruvate delivery to the tumor cells and is less affected by the endogenous lactate concentration.
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- 2021
9. A Critical and Concise Review of Mass Spectrometry Applied to Imaging in Drug Discovery
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Josephine Bunch, Zoltan Takats, Richard J. A. Goodwin, INSERM, Université de Lille, Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM) - U1192, University of Glasgow, Imperial College London, Protéomique, Réponse Inflammatoire, Spectrométrie de Masse (PRISM) - U 1192 (PRISM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)
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Drug ,Biodistribution ,MALDI ,imaging ,MSI ,mass spectrometry imaging ,SIMS ,DESI ,media_common.quotation_subject ,[SDV]Life Sciences [q-bio] ,Computational biology ,Mass spectrometry ,01 natural sciences ,Biochemistry ,Mass spectrometry imaging ,Analytical Chemistry ,03 medical and health sciences ,Pharmacokinetics ,Drug Discovery ,Image Processing, Computer-Assisted ,Medicine ,Humans ,Tissue Distribution ,Pharmaceutical sciences ,030304 developmental biology ,media_common ,0303 health sciences ,Drug discovery ,business.industry ,010401 analytical chemistry ,0104 chemical sciences ,Molecular Imaging ,Tissue sections ,Pharmaceutical Preparations ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Molecular Medicine ,business ,Biotechnology - Abstract
International audience; During the past decade, mass spectrometry imaging (MSI) has become a robust and versatile methodology to support modern pharmaceutical research and development. The technologies provide data on the biodistribution, metabolism, and delivery of drugs in tissues, while also providing molecular maps of endogenous metabolites, lipids, and proteins. This allows researchers to make both pharmacokinetic and pharmacodynamic measurements at cellular resolution in tissue sections or clinical biopsies. Despite drug imaging within samples now playing a vital role within research and development (R&D) in leading pharmaceutical companies, however, the challenges in turning compounds into medicines continue to evolve as rapidly as the technologies used to discover them. The increasing cost of development of new and emerging therapeutic modalities, along with the associated risks of late-stage program attrition, means there is still an unmet need in our ability to address an increasing array of challenging bioanalytical questions within drug discovery. We require new capabilities and strategies of integrated imaging to provide context for fundamental disease-related biological questions that can also offer insights into specific project challenges. Integrated molecular imaging and advanced image analysis have the opportunity to provide a world-class capability that can be deployed on projects in which we cannot answer the question with our battery of established assays. Therefore, here we will provide an updated concise review of the use of MSI for drug discovery; we will also critically consider what is required to embed MSI into a wider evolving R&D landscape and allow long-lasting impact in the industry.
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- 2020
10. Universal Sample Preparation Unlocking Multimodal Molecular Tissue Imaging
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Owen J. Sansom, Nicole Strittmatter, John G. Swales, Gregory Hamm, Stephanie Ling, Andreas Dannhorn, Chelsea J. Nikula, Simon T. Barry, Paolo Inglese, Emine Kazanc, Jean-Luc Vorng, Evdoxia Karali, Maria Paola Serra, George Poulogiannis, Gareth Maglennon, Zoltan Takats, Richard J. A. Goodwin, and Josephine Bunch
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Male ,Spectrometry, Mass, Electrospray Ionization ,Tissue imaging ,Electrospray ionization ,010402 general chemistry ,Mass spectrometry ,Real-Time Polymerase Chain Reaction ,01 natural sciences ,Analytical Chemistry ,Specimen Handling ,Hypromellose Derivatives ,medicine ,Animals ,Sample preparation ,Rats, Wistar ,Desorption electrospray ionization ,Polyvinylpyrrolidone ,Tissue Embedding ,Chemistry ,010401 analytical chemistry ,Povidone ,Reproducibility of Results ,Hydrogels ,0104 chemical sciences ,Secondary ion mass spectrometry ,Matrix-assisted laser desorption/ionization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,medicine.drug ,Biomedical engineering - Abstract
A new tissue sample embedding and processing method is presented that provides downstream compatibility with numerous different histological, molecular biology, and analytical techniques. The methodology is based on the low temperature embedding of fresh frozen specimens into a hydrogel matrix composed of hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP) and sectioning using a cryomicrotome. The hydrogel was expected not to interfere with standard tissue characterization methods, histologically or analytically. We assessed the compatibility of this protocol with various mass spectrometric imaging methods including matrix-assisted laser desorption ionization (MALDI), desorption electrospray ionization (DESI) and secondary ion mass spectrometry (SIMS). We also demonstrated the suitability of the universal protocol for extraction based molecular biology techniques such as rt-PCR. The integration of multiple analytical modalities through this universal sample preparation protocol offers the ability to study tissues at a systems biology level and directly linking results to tissue morphology and cellular phenotype.
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- 2020
11. LESA MS Imaging of Heat-Preserved and Frozen Tissue: Benefits of Multistep Static FAIMS
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Richard J. A. Goodwin, Anna L. Simmonds, Helen J. Cooper, Rian L. Griffiths, and John G. Swales
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Male ,0301 basic medicine ,Hot Temperature ,Surface Properties ,Ion-mobility spectrometry ,Liquid-Liquid Extraction ,Single step ,Kidney ,Mass spectrometry ,01 natural sciences ,Mass spectrometry imaging ,Analytical Chemistry ,03 medical and health sciences ,Tandem Mass Spectrometry ,Ion Mobility Spectrometry ,Testis ,Animals ,Rats, Wistar ,Frozen tissue ,Brain Chemistry ,Chromatography ,Chemistry ,010401 analytical chemistry ,Proteins ,0104 chemical sciences ,Cold Temperature ,030104 developmental biology ,Tissue sections ,High field ,Asymmetric waveform - Abstract
We have previously demonstrated liquid extraction surface analysis (LESA) high field asymmetric waveform ion mobility spectrometry (FAIMS) mass spectrometry imaging of proteins in thin tissue sections of brain and liver. Here, we present an improved approach that makes use of multiple static FAIMS parameters at each sampled location and allows a significant improvement in the number of proteins, lipids, and drugs that can be imaged simultaneously. The approach is applied to the mass spectrometry imaging of control and cassette-dosed rat kidneys. Mass spectrometry imaging of kidneys typically requires washing to remove excess hemoglobin; however, that is not necessary with this approach. Multistep static FAIMS mass spectrometry resulted in a 6- to 16-fold increase in the number of proteins detected in the absence of FAIMS, in addition to smaller increases over single step static FAIMS (chosen for optimum transmission of total protein ions). The benefits of multistep static FAIMS mass spectrometry for protein detection are also shown for sections of testes. The numbers of proteins detected following multistep FAIMS increased between 2- and 3-fold over single step FAIMS and between 2- and 14-fold over LESA alone. Finally, to date, LESA mass spectrometry of proteins in tissue has been undertaken solely on fresh frozen samples. In this work, we demonstrate that heat-preserved tissues are also suitable for these analyses. Heat preservation of tissue improved the number of proteins detected by LESA MS for both kidney and testes tissue (by between 2- and 4-fold). For both tissue types, the majority of the proteins additionally detected in the heat-treated samples were subsequently detected in the frozen samples when FAIMS was incorporated. Improvements in the numbers of proteins detected were observed for LESA FAIMS MS for the kidney tissue; for testes tissue, fewer total proteins were detected following heat preservation; however, approximately one-third were unique to the heat-preserved samples.
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- 2018
12. Nanoextraction coupled to liquid chromatography mass spectrometry delivers improved spatially resolved analysis
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Janella de Jesus, Holly-May Lewis, Vladimir Palitsin, Richard J. A. Goodwin, Josephine Bunch, Melanie J. Bailey, Roger P. Webb, John G. Swales, Patrick Sears, Guido F. Verbeck, and Catia Costa
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Analyte ,Chromatography ,Chemistry ,Capillary action ,010401 analytical chemistry ,Extraction (chemistry) ,Repeatability ,010402 general chemistry ,Mass spectrometry ,01 natural sciences ,0104 chemical sciences ,Analytical Chemistry ,Solvent ,Matrix (chemical analysis) ,Liquid chromatography–mass spectrometry - Abstract
Direct analyte probed nanoextraction (DAPNe) is a technique that allows extraction of drug and endogenous compounds from a discrete location on a tissue sample using a nano capillary filled with solvent. Samples can be extracted from a spot diameters as low as 6 µm. Studies previously undertaken by our group have shown that the technique can provide good precision (5%) for analysing drug molecules in 150 µm diameter areas of homogenised tissue, provided an internal standard is sprayed on to the tissue prior to analysis. However, without an isotopically labelled standard, the repeatability is poor, even after normalisation to and the spot area or matrix compounds. By application to tissue homogenates spiked with drug compounds, we can demonstrate that it is possible to significantly improve the repeatability of the technique by incorporating a liquid chromatography separation step. Liquid chromatography is a technique for separating compounds prior to mass spectrometry (LC-MS) which enables separation of isomeric compounds that cannot be discriminated using mass spectrometry alone, as well as reducing matrix interferences. Conventionally, LC-MS is carried out on bulk or homogenised samples, which means analysis is essentially an average of the sample and does not take into account discrete areas. This work opens a new opportunity for spatially resolved liquid chromatography mass spectrometry with precision better than 20%.
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- 2019
13. Quantitation of Endogenous Metabolites in Mouse Tumors Using Mass-Spectrometry Imaging
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Malcolm R. Clench, Pablo Morentin-Gutierrez, Nicole Strittmatter, John G. Swales, Filippos Michopoulos, Christopher Hardy, Martine J. Mellor, Gregory Hamm, Alex Dexter, Anna Nilsson, Richard J. A. Goodwin, Josephine Bunch, Per E. Andrén, and Susan E. Critchlow
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0301 basic medicine ,Analyte ,Reproducibility ,Chromatography ,Chemistry ,010401 analytical chemistry ,Glutamic Acid ,Mice, Nude ,Endogeny ,Neoplasms, Experimental ,Mass spectrometry ,01 natural sciences ,Mass Spectrometry ,Mass spectrometry imaging ,0104 chemical sciences ,Analytical Chemistry ,Mice ,03 medical and health sciences ,030104 developmental biology ,Tissue sections ,Animals ,Female ,Mouse tumor ,Lactic Acid ,Glutamic acid metabolism - Abstract
Described is a quantitative-mass-spectrometryimaging\ud (qMSI) methodology for the analysis of lactate and\ud glutamate distributions in order to delineate heterogeneity\ud among mouse tumor models used to support drug-discovery\ud efficacy testing. We evaluate and report on preanalysisstabilization\ud methods aimed at improving the reproducibility\ud and efficiency of quantitative assessments of endogenous\ud molecules in tissues. Stability experiments demonstrate that\ud optimum stabilization protocols consist of frozen-tissue\ud embedding, post-tissue-sectioning desiccation, and storage at\ud −80 °C of tissue sections sealed in vacuum-tight containers.\ud Optimized stabilization protocols are used in combination with qMSI methodology for the absolute quantitation of lactate and\ud glutamate in tumors, incorporating the use of two different stable-isotope-labeled versions of each analyte and spectral-clustering\ud performed on each tissue section using k-means clustering to allow region-specific, pixel-by-pixel quantitation. Region-specific\ud qMSI was used to screen different tumor models and identify a phenotype that has low lactate heterogeneity, which will enable\ud accurate measurements of lactate modulation in future drug-discovery studies. We conclude that using optimized qMSI\ud protocols, it is possible to quantify endogenous metabolites within tumors, and region-specific quantitation can provide valuable\ud insight into tissue heterogeneity and the tumor microenvironment.
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- 2018
14. Uncovering the regional localization of inhaled salmeterol retention in the lung
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Gregory Hamm, Britt-Marie Fihn, Erica Bäckström, Anna Nilsson, Richard J. A. Goodwin, Per E. Andrén, Markus Fridén, and Nicole Strittmatter
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Male ,0301 basic medicine ,lung retention ,Respiratory Medicine and Allergy ,Polysorbates ,Pharmaceutical Science ,Pharmacology ,Mass Spectrometry ,Polyethylene Glycols ,Cluster Analysis ,Tissue Distribution ,Lung ,Salmeterol Xinafoate ,Lungmedicin och allergi ,media_common ,inhalation ,Inhalation ,Respiratory disease ,General Medicine ,respiratory system ,Farmakologi och toxikologi ,pulmonary distribution ,Bronchodilator Agents ,Molecular Imaging ,medicine.anatomical_structure ,Injections, Intravenous ,Systemic administration ,Original Article ,Salmeterol ,Pharmaceutical Vehicles ,pharmacokinetics ,medicine.drug ,Drug ,media_common.quotation_subject ,Bronchi ,Respiratory Mucosa ,Pharmacology and Toxicology ,mass spectrometry imaging ,03 medical and health sciences ,Pharmacokinetics ,Administration, Inhalation ,medicine ,Animals ,Distribution (pharmacology) ,Pulmonary distribution ,Rats, Wistar ,Adrenergic beta-2 Receptor Agonists ,business.industry ,Phosphatidylethanolamines ,lcsh:RM1-950 ,Deuterium ,medicine.disease ,respiratory tract diseases ,Pulmonary Alveoli ,Respiratory Tract Absorption ,lcsh:Therapeutics. Pharmacology ,030104 developmental biology ,business - Abstract
Treatment of respiratory disease with a drug delivered via inhalation is generally held as being beneficial as it provides direct access to the lung target site with a minimum systemic exposure. There is however only limited information of the regional localization of drug retention following inhalation. The aim of this study was to investigate the regional and histological localization of salmeterol retention in the lungs after inhalation and to compare it to systemic administration. Lung distribution of salmeterol delivered to rats via nebulization or intravenous (IV) injection was analyzed with high-resolution mass spectrometry imaging (MSI). Salmeterol was widely distributed in the entire section at 5 min after inhalation, by 15 min it was preferentially retained in bronchial tissue. Via a novel dual-isotope study, where salmeterol was delivered via inhalation and d3-salmeterol via IV to the same rat, could the effective gain in drug concentration associated with inhaled delivery relative to IV, expressed as a site-specific lung targeting factor, was 5-, 31-, and 45-fold for the alveolar region, bronchial sub-epithelium and epithelium, respectively. We anticipate that this MSI-based framework for quantifying regional and histological lung targeting by inhalation will accelerate discovery and development of local and more precise treatments of respiratory disease.
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- 2018
15. Quantitative Imaging of Proteins in Tissue by Stable Isotope Labeled Mimetic Liquid Extraction Surface Analysis Mass Spectrometry
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Elizabeth C. Randall, John G. Swales, Iain B. Styles, Rian L. Griffiths, Helen J. Cooper, Richard J. A. Goodwin, Josephine Bunch, and Jana Havlikova
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Quantitative imaging ,Letter ,Liquid-Liquid Extraction ,Endogeny ,010402 general chemistry ,Mass spectrometry ,01 natural sciences ,Mass Spectrometry ,Analytical Chemistry ,Mice ,Ubiquitin ,Animals ,Sample preparation ,Brain Chemistry ,Chromatography ,biology ,Stable isotope ratio ,Chemistry ,Spatially resolved ,010401 analytical chemistry ,0104 chemical sciences ,Rats ,biology.protein ,Homogenization (biology) ,Chromatography, Liquid - Abstract
Absolute quantification of proteins in tissue is important for numerous fields of study. Liquid chromatography–mass spectrometry (LC–MS) methods are the norm but typically involve lengthy sample preparation including tissue homogenization, which results in the loss of information relating to spatial distribution. Here, we propose liquid extraction surface analysis (LESA) mass spectrometry (MS) of stable isotope labeled mimetic tissue models for the spatially resolved quantification of intact ubiquitin in rat and mouse brain tissue. Measured ubiquitin concentrations are in agreement with values found in the literature. Images of rat and mouse brain tissue demonstrate spatial variation in the concentration of ubiquitin and demonstrate the utility of spatially resolved quantitative measurement of proteins in tissue. Although we have focused on ubiquitin, the method has the potential for broader application to the absolute quantitation of any endogenous protein or protein-based drug in tissue.
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- 2019
16. Liquid Extraction Surface Analysis (LESA) Electron-Induced Dissociation and Collision-Induced Dissociation Mass Spectrometry of Small Molecule Drug Compounds
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Helen J. Cooper, Rian L. Griffiths, Richard J. A. Goodwin, Andrea F. Lopez-Clavijo, Cooper, Helen J [0000-0003-4590-9384], and Apollo - University of Cambridge Repository
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Male ,Electrospray ,Collision-induced dissociation ,Surface Properties ,Liquid-Liquid Extraction ,010402 general chemistry ,Mass spectrometry ,Tandem mass spectrometry ,Kidney ,01 natural sciences ,Electron ionisation ,Dissociation (chemistry) ,Liquid extraction surface analysis ,CID ,Small molecules dissociation ,EID ,Structural Biology ,Tandem Mass Spectrometry ,Drug compounds ,Animals ,Pharmacokinetics ,Rats, Wistar ,Spectroscopy ,Electron ionization ,Chromatography ,Electron-induced dissociation ,Chemistry ,LESA ,010401 analytical chemistry ,EI ,0104 chemical sciences ,Rats ,Pharmaceutical Preparations ,Mass spectrum ,Ion cyclotron resonance - Abstract
Here, we present liquid extraction surface analysis (LESA) coupled with electron-induced dissociation (EID) mass spectrometry in a Fourier-transform ion cyclotron resonance mass spectrometer for the analysis of small organic pharmaceutical compounds directly from dosed tissue. First, the direct infusion electrospray ionisation EID and collision-induced dissociation (CID) behaviour of erlotinib, moxifloxacin, clozapine and olanzapine standards were compared. EID mass spectra were also compared with experimental or reference electron impact ionisation mass spectra. The results show that (with the exception of erlotinib) EID and CID result in complementary fragment ions. Subsequently, we performed LESA EID MS/MS and LESA CID MS/MS on singly charged ions of moxifloxacin and erlotinib extracted from a thin tissue section of rat kidney from a cassette-dosed animal. Both techniques provided structural information, with the majority of peaks observed for the drug standards also observed for the tissue-extracted species. Overall, these results demonstrate the feasibility of LESA EID MS/MS of drug compounds from dosed tissue and extend the number of molecular structures for which EID behaviour has been determined. Graphical Abstract ᅟ.
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- 2018
17. Exemplifying the Screening Power of Mass Spectrometry Imaging over Label-Based Technologies for Simultaneous Monitoring of Drug and Metabolite Distributions in Tissue Sections
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Anna Nilsson, Richard J. A. Goodwin, Martin Billger, John G. Swales, Per E. Andrén, Maria Johansson, Suzanne L. Iverson, and C. Logan Mackay
- Subjects
Diagnostic Imaging ,Male ,0301 basic medicine ,Metabolite ,Pharmacology ,Mass spectrometry ,01 natural sciences ,Biochemistry ,Mass spectrometry imaging ,Fourier transform ion cyclotron resonance ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Neoplasms ,Drug Discovery ,Animals ,Distribution (pharmacology) ,Chromatography ,Drug discovery ,Chemistry ,010401 analytical chemistry ,0104 chemical sciences ,Mice, Inbred C57BL ,Matrix-assisted laser desorption/ionization ,030104 developmental biology ,Pharmaceutical Preparations ,Blood-Brain Barrier ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Autoradiography ,Molecular Medicine ,Peptides ,Drug metabolism ,Biotechnology - Abstract
Mass spectrometry imaging (MSI) provides pharmaceutical researchers with a suite of technologies to screen and assess compound distributions and relative abundances directly from tissue sections and offer insight into drug discovery-applicable queries such as blood-brain barrier access, tumor penetration/retention, and compound toxicity related to drug retention in specific organs/cell types. Label-free MSI offers advantages over label-based assays, such as quantitative whole-body autoradiography (QWBA), in the ability to simultaneously differentiate and monitor both drug and drug metabolites. Such discrimination is not possible by label-based assays if a drug metabolite still contains the radiolabel. Here, we present data exemplifying the advantages of MSI analysis. Data of the distribution of AZD2820, a therapeutic cyclic peptide, are related to corresponding QWBA data. Distribution of AZD2820 and two metabolites is achieved by MSI, which [(14)C]AZD2820 QWBA fails to differentiate. Furthermore, the high mass-resolving power of Fourier transform ion cyclotron resonance MS is used to separate closely associated ions.
- Published
- 2016
18. Application of Various Normalization Methods for Microscale Analysis of Tissues Using Direct Analyte Probed Nanoextraction
- Author
-
Janella de Jesus, Catia Costa, Roger P. Webb, Richard J. A. Goodwin, Josephine Bunch, Guido F. Verbeck, and Melanie J. Bailey
- Subjects
0301 basic medicine ,Normalization (statistics) ,Organelles ,Analyte ,Chemistry ,Sample (material) ,010401 analytical chemistry ,Extraction (chemistry) ,Replicate ,Mass spectrometry ,01 natural sciences ,Mass Spectrometry ,0104 chemical sciences ,Analytical Chemistry ,03 medical and health sciences ,Matrix (mathematics) ,030104 developmental biology ,Liver ,Humans ,Nanotechnology ,Single-Cell Analysis ,Biological system ,Microscale chemistry - Abstract
Direct analyte-probed nano-extraction (DAPNe) is a method of extracting material from a microscale region of a sample and provides the opportunity for detailed mass spectrometry analysis of extracted analytes from a small area. The technique has been shown to provide enhanced sensitivity compared with bulk analysis by selectively removing analytes from their matrix and has been applied for selective analysis of single cells and even single organelles. However, the quantitative capabilities of the technique are yet to be fully evaluated. In this study, various normalisation techniques were investigated in order to improve the quantitative capabilities of the technique. Two methods of internal standard incorporation were applied to test substrates, which were designed to replicate biological sample matrices. Additionally, normalisation to the extraction spot area and matrix compounds were investigated for suitability in situations when an internal standard is not available. The variability observed can be significantly reduced by using a sprayed internal standard, and in some cases, by normalising to the extracted area.
- Published
- 2018
19. Spatial Quantitation of Drugs in tissues using Liquid Extraction Surface Analysis Mass Spectrometry Imaging
- Author
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Peter J. H. Webborn, Malcolm R. Clench, Nicole Strittmatter, John G. Swales, James W. Tucker, and Richard J. A. Goodwin
- Subjects
0301 basic medicine ,Male ,Analyte ,Spectrometry, Mass, Electrospray Ionization ,Calibration curve ,Electrospray ionization ,Tandem mass spectrometry ,Mass spectrometry ,01 natural sciences ,Mass spectrometry imaging ,Article ,03 medical and health sciences ,Imaging, Three-Dimensional ,Limit of Detection ,Tandem Mass Spectrometry ,Animals ,Rats, Wistar ,Detection limit ,Multidisciplinary ,Chromatography ,Chemistry ,010401 analytical chemistry ,Analytical technique ,Reference Standards ,0104 chemical sciences ,030104 developmental biology ,Pharmaceutical Preparations ,Organ Specificity ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Calibration ,Chromatography, Liquid - Abstract
Liquid extraction surface analysis mass spectrometry imaging (LESA-MSI) has been shown to be an effective tissue profiling and imaging technique, producing robust and reliable qualitative distribution images of an analyte or analytes in tissue sections. Here, we expand the use of LESA-MSI beyond qualitative analysis to a quantitative analytical technique by employing a mimetic tissue model previously shown to be applicable for MALDI-MSI quantitation. Liver homogenate was used to generate a viable and molecularly relevant control matrix for spiked drug standards which can be frozen, sectioned and subsequently analyzed for the generation of calibration curves to quantify unknown tissue section samples. The effects of extraction solvent composition, tissue thickness and solvent/tissue contact time were explored prior to any quantitative studies in order to optimize the LESA-MSI method across several different chemical entities. The use of a internal standard to normalize regional differences in ionization response across tissue sections was also investigated. Data are presented comparing quantitative results generated by LESA-MSI to LC-MS/MS. Subsequent analysis of adjacent tissue sections using DESI-MSI is also reported.
- Published
- 2016
20. Conductive carbon tape used for support and mounting of both whole animal and fragile heat-treated tissue sections for MALDI MS imaging and quantitation
- Author
-
David J. Harrison, Anna Nilsson, Per E. Andrén, Suzanne L. Iverson, C. Logan Mackay, Patrick R. R. Langridge-Smith, Richard J. A. Goodwin, and Daniel Borg
- Subjects
Diagnostic Imaging ,Male ,Restraint, Physical ,Hot Temperature ,Maldi ms ,Materials science ,Biophysics ,Nanotechnology ,Mass spectrometry ,Biochemistry ,Mass spectrometry imaging ,Specimen Handling ,Animals ,Whole Body Imaging ,Surgical Tape ,Paraffin Embedding ,Histological Techniques ,Thermal Conductivity ,Microtomy ,Carbon ,Heat stabilization ,Rats ,Tissue sections ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Heat treated ,Sample collection ,WHOLE ANIMAL ,Biomedical engineering - Abstract
Analysis of whole animal tissue sections by MALDI MS imaging (MSI) requires effective sample collection and transfer methods to allow the highest quality of in situ analysis of small or hard to dissect tissues. We report on the use of double-sided adhesive conductive carbon tape during whole adult rat tissue sectioning of carboxymethyl cellulose (CMC) embedded animals, with samples mounted onto large format conductive glass and conductive plastic MALDI targets, enabling MSI analysis to be performed on both TOF and FT-ICR MALDI mass spectrometers. We show that mounting does not unduly affect small molecule MSI detection by analyzing tiotropium abundance and distribution in rat lung tissues, with direct on-tissue quantitation achieved. Significantly, we use the adhesive tape to provide support to embedded delicate heat-stabilized tissues, enabling sectioning and mounting to be performed that maintained tissue integrity on samples that had previously been impossible to adequately prepare section for MSI analysis. The mapping of larger peptidomic molecules was not hindered by tape mounting samples and we demonstrate this by mapping the distribution of PEP-19 in both native and heat-stabilized rat brains. Furthermore, we show that without heat stabilization PEP-19 degradation fragments can detected and identified directly by MALDI MSI analysis. This article is part of a Special Issue entitled: Imaging Mass Spectrometry: A User’s Guide to a New Technique for Biological and Biomedical Research.
- Published
- 2012
21. Deuterated Matrix-Assisted Laser Desorption Ionization Matrix Uncovers Masked Mass Spectrometry Imaging Signals of Small Molecules
- Author
-
Per Svenningsson, Richard J. A. Goodwin, Laszlo Kladni, Zoltan Banka, Anna Nilsson, Nicoletta Schintu, Mohammadreza Shariatgorji, Per E. Andrén, Tibor Hasko, and Andras Szabo
- Subjects
Male ,Berberine ,Coumaric Acids ,Analytical chemistry ,Brain ,Mass spectrometry ,Small molecule ,Acetylcholine ,Mass spectrometry imaging ,Molecular Imaging ,Rats ,Analytical Chemistry ,Molecular Weight ,Matrix (chemical analysis) ,Mice ,chemistry.chemical_compound ,Matrix-assisted laser desorption/ionization ,chemistry ,Deuterium ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Desorption ,Animals ,Isoquinoline ,Lung - Abstract
D(4)-α-Cyano-4-hydroxycinnamic acid (D(4)-CHCA) has been synthesized for use as a matrix for matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) and MALDI-MS imaging (MSI) of small molecule drugs and endogenous compounds. MALDI-MS analysis of small molecules has historically been hindered by interference from matrix ion clusters and fragment peaks that mask signals of low molecular weight compounds of interest. By using D(4)-CHCA, the cluster and fragment peaks of CHCA, the most common matrix for analysis of small molecules, are shifted by + 4, + 8 and + 12 Da, which expose signals across areas of the previously concealed low mass range. Here, obscured MALDI-MS signals of a synthetic small molecule pharmaceutical, a naturally occurring isoquinoline alkaloid, and endogenous compounds including the neurotransmitter acetylcholine have been unmasked and imaged directly from biological tissue sections.
- Published
- 2012
22. Simultaneous imaging of multiple neurotransmitters and neuroactive substances in the brain by desorption electrospray ionization mass spectrometry
- Author
-
Xiaoqun Zhang, Mohammadreza Shariatgorji, Per Svenningsson, Anna Nilsson, Theodosia Vallianatou, Alexandra Alvarsson, Per E. Andrén, Patrik Källback, Richard J. A. Goodwin, and Nicole Strittmatter
- Subjects
0301 basic medicine ,Male ,Serotonin ,Spectrometry, Mass, Electrospray Ionization ,Neurologi ,Dopamine ,Parkinson's disease ,Cognitive Neuroscience ,Mass spectrometry ,01 natural sciences ,Sensitivity and Specificity ,Imaging ,Rats, Sprague-Dawley ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,medicine ,Animals ,Neurochemistry ,Tissue Distribution ,Neurotransmitter ,Amphetamine ,Neurotransmitter Agents ,Psychotropic Drugs ,Chromatography ,Drug discovery ,010401 analytical chemistry ,Brain ,Reproducibility of Results ,Signal Processing, Computer-Assisted ,Magnetic Resonance Imaging ,0104 chemical sciences ,3. Good health ,Molecular Imaging ,Rats ,Glutamine ,Mice, Inbred C57BL ,030104 developmental biology ,Neurology ,chemistry ,Radiologi och bildbehandling ,Algorithms ,Radiology, Nuclear Medicine and Medical Imaging ,medicine.drug - Abstract
With neurological processes involving multiple neurotransmitters and neuromodulators, it is important to have the ability to directly map and quantify multiple signaling molecules simultaneously in a single analysis. By utilizing a molecular-specific approach, namely desorption electrospray ionization mass spectrometry imaging (DESI-MSI), we demonstrated that the technique can be used to image multiple neurotransmitters and their metabolites (dopamine, dihydroxyphenylacetic acid, 3-methoxytyramine, serotonin, glutamate, glutamine, aspartate,gamma-aminobutyric acid, adenosine) as well as neuroactive drugs (amphetamine, sibutramine, fluvoxamine) and drug metabolites in situ directly in brain tissue sections. The use of both positive and negative ionization modes increased the number of identified molecular targets. Chemical derivatization by charge-tagging the primary amines of molecules significantly increased the sensitivity, enabling the detection of low abundant neurotransmitters and other neuroactive substances previously undetectable by MSI. The sensitivity of the imaging approach of neurochemicals has a great potential in many diverse applications in fields such as neuroscience, pharmacology, drug discovery, neurochemistry, and medicine.
- Published
- 2015
23. Qualitative and Quantitative MALDI Imaging of the Positron Emission Tomography Ligands Raclopride (a D2 Dopamine Antagonist) and SCH 23390 (a D1 Dopamine Antagonist) in Rat Brain Tissue Sections Using a Solvent-Free Dry Matrix Application Method
- Author
-
Richard J. A. Goodwin, C. Logan Mackay, Lars Farde, David J. Harrison, Anna Nilsson, Suzanne L. Iverson, and Per E. Andrén
- Subjects
MALDI imaging ,Ligands ,Mass spectrometry ,Analytical Chemistry ,Rats, Sprague-Dawley ,Matrix (chemical analysis) ,chemistry.chemical_compound ,Nuclear magnetic resonance ,Desorption ,medicine ,Animals ,Raclopride ,SCH-23390 ,Chromatography ,medicine.diagnostic_test ,Dopamine antagonist ,Brain ,Tin Compounds ,Benzazepines ,Rats ,chemistry ,Positron emission tomography ,Positron-Emission Tomography ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Dopamine Antagonists ,medicine.drug - Abstract
The distributions of positron emission tomography (PET) ligands in rat brain tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI). The detection of the PET ligands was possible following the use of a solvent-free dry MALDI matrix application method employing finely ground dry α-cyano-4-hydroxycinnamic acid (CHCA). The D2 dopamine receptor antagonist 3,5-dichloro-N-{[(2S)-1-ethylpyrrolidin-2-yl]methyl}-2-hydroxy-6-methoxybenzamide (raclopride) and the D1 dopamine receptor antagonist 7-chloro-3-methyl-1-phenyl-1,2,4,5-tetrahydro-3-benzazepin-8-ol (SCH 23390) were both detected at decreasing abundance at increasing period postdosing. Confirmation of the compound identifications and distributions was achieved by a combination of mass-to-charge ratio accurate mass, isotope distribution, and MS/MS fragmentation imaging directly from tissue sections (performed using MALDI TOF/TOF, MALDI q-TOF, and 12T MALDI-FT-ICR mass spectrometers). Quantitative data was obtained by comparing signal abundances from tissues to those obtained from quantitation control spots of the target compound applied to adjacent vehicle control tissue sections (analyzed during the same experiment). Following a single intravenous dose of raclopride (7.5 mg/kg), an average tissue concentration of approximately 60 nM was detected compared to 15 nM when the drug was dosed at 2 mg/kg, indicating a linear response between dose and detected abundance. SCH 23390 was established to have an average tissue concentration of approximately 15 μM following a single intravenous dose at 5 mg/kg. Both target compounds were also detected in kidney tissue sections when employing the same MSI methodology. This study illustrates that a MSI may well be readily applied to PET ligand research development when using a solvent-free dry matrix coating.
- Published
- 2011
24. Matrix‐free mass spectrometric imaging using laser desorption ionisation <scp>F</scp> ourier transform ion cyclotron resonance mass spectrometry
- Author
-
David J. Harrison, Stefan Weidt, C. Logan Mackay, Patrick R. R. Langridge-Smith, Andrew R. Pitt, Richard J. A. Goodwin, and Michael P. Barrett
- Subjects
Brain Chemistry ,MALDI imaging ,Chromatography ,Fourier Analysis ,Protein mass spectrometry ,Histocytochemistry ,Chemistry ,Organic Chemistry ,Analytical chemistry ,Tin Compounds ,Kidney ,Mass spectrometry ,Fourier transform ion cyclotron resonance ,Sample preparation in mass spectrometry ,Mass spectrometry imaging ,Analytical Chemistry ,Surface-enhanced laser desorption/ionization ,Mice ,Matrix-assisted laser desorption/ionization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Animals ,Metabolomics ,Phospholipids ,Research Articles ,Spectroscopy - Abstract
Mass spectrometry imaging (MSI) is a powerful tool in metabolomics and proteomics for the spatial localization and identification of pharmaceuticals, metabolites, lipids, peptides and proteins in biological tissues. However, sample preparation remains a crucial variable in obtaining the most accurate distributions. Common washing steps used to remove salts, and solvent-based matrix application, allow analyte spreading to occur. Solvent-free matrix applications can reduce this risk, but increase the possibility of ionisation bias due to matrix adhesion to tissue sections. We report here the use of matrix-free MSI using laser desorption ionisation performed on a 12 T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. We used unprocessed tissue with no post-processing following thaw-mounting on matrix-assisted laser desorption ionisation (MALDI) indium-tin oxide (ITO) target plates. The identification and distribution of a range of phospholipids in mouse brain and kidney sections are presented and compared with previously published MALDI time-of-flight (TOF) MSI distributions. Copyright © 2011 John Wiley & Sons, Ltd.
- Published
- 2011
25. A solvent-free matrix application method for matrix-assisted laser desorption/ionization imaging of small molecules
- Author
-
S.P. Scullion, Andrew R. Pitt, Richard J. A. Goodwin, David G. Watson, and Lynsey MacIntyre
- Subjects
Matrix (chemical analysis) ,Analyte ,Matrix-assisted laser desorption/ionization ,Chemistry ,Ionization ,Desorption ,Organic Chemistry ,Analytical chemistry ,Sample preparation ,Mass spectrometry ,Spectroscopy ,Mass spectrometry imaging ,Analytical Chemistry - Abstract
Matrix application continues to be a critical step in sample preparation for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI). Imaging of small molecules such as drugs and metabolites is particularly problematic because the commonly used washing steps to remove salts are usually omitted as they may also remove the analyte, and analyte spreading is more likely with conventional wet matrix application methods. We have developed a method which uses the application of matrix as a dry, finely divided powder, here referred to as dry matrix application, for the imaging of drug compounds. This appears to offer a complementary method to wet matrix application for the MALDI-MSI of small molecules, with the alternative matrix application techniques producing different ion profiles, and allows the visualization of compounds not observed using wet matrix application methods. We demonstrate its value in imaging clozapine from rat kidney and 4-bromophenyl-1,4-diazabicyclo(3.2.2)nonane-4-carboxylic acid from rat brain. In addition, exposure of the dry matrix coated sample to a saturated moist atmosphere appears to enhance the visualization of a different set of molecules.
- Published
- 2010
26. Future directions of imaging MS in pharmaceutical RD
- Author
-
Peter J. H. Webborn and Richard J. A. Goodwin
- Subjects
Engineering ,Drug Industry ,Management science ,business.industry ,Research areas ,Drug discovery ,Research ,Clinical Biochemistry ,Nanotechnology ,General Medicine ,Mass Spectrometry ,Analytical Chemistry ,Medical Laboratory Technology ,Pharmaceutical Preparations ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Drug Discovery ,Humans ,General Pharmacology, Toxicology and Pharmaceutics ,Pharmaceutical sciences ,business ,Drug industry - Abstract
MS imaging has rapidly evolved over the last decade, finding roles in all aspects of pharmaceutical research and development. This article discusses possible methodological and technological future advancements and describes research areas where the technology can expand and continue to prove to be worthwhile tool for drug discovery and development.
- Published
- 2015
27. Mapping drug distribution in brain tissue using liquid extraction surface analysis mass spectrometry imaging
- Author
-
Peter J. H. Webborn, Richard J. A. Goodwin, Malcolm R. Clench, James W. Tucker, John G. Swales, Suzanne L. Iverson, and Michael J. Spreadborough
- Subjects
MALDI imaging ,Male ,Spectrometry, Mass, Electrospray Ionization ,Chromatography ,Chemistry ,Blood contamination ,Brain ,Brain tissue ,Mass spectrometry ,Mass spectrometry imaging ,Analytical Chemistry ,Rats ,Pharmaceutical Preparations ,Liquid chromatography–mass spectrometry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Distribution (pharmacology) ,Animals ,Pharmacokinetics ,Tissue Distribution ,Rats, Wistar ,Image resolution - Abstract
Liquid extraction surface analysis mass spectrometry (LESA-MS) is a surface sampling technique that incorporates liquid extraction from the surface of tissue sections with nanoelectrospray mass spectrometry. Traditional tissue analysis techniques usually require homogenization of the sample prior to analysis via high-performance liquid chromatography mass spectrometry (HPLC-MS), but an intrinsic weakness of this is a loss of all spatial information and the inability of the technique to distinguish between actual tissue penetration and response caused by residual blood contamination. LESA-MS, in contrast, has the ability to spatially resolve drug distributions and has historically been used to profile discrete spots on the surface of tissue sections. Here, we use the technique as a mass spectrometry imaging (MSI) tool, extracting points at 1 mm spatial resolution across tissue sections to build an image of xenobiotic and endogenous compound distribution to assess drug blood-brain barrier penetration into brain tissue. A selection of penetrant and "nonpenetrant" drugs were dosed to rats via oral and intravenous administration. Whole brains were snap-frozen at necropsy and were subsequently sectioned prior to analysis by matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and LESA-MSI. MALDI-MSI, as expected, was shown to effectively map the distribution of brain penetrative compounds but lacked sufficient sensitivity when compounds were marginally penetrative. LESA-MSI was used to effectively map the distribution of these poorly penetrative compounds, highlighting its value as a complementary technique to MALDI-MSI. The technique also showed benefits when compared to traditional homogenization, particularly for drugs that were considered nonpenetrant by homogenization but were shown to have a measurable penetration using LESA-MSI.
- Published
- 2015
28. Mass spectrometry imaging in drug development
- Author
-
Anna Nilsson, Mohammadreza Shariatgorji, Per E. Andrén, Peter J. H. Webborn, Theodosia Vallianatou, and Richard J. A. Goodwin
- Subjects
Chromatography ,Drug-Related Side Effects and Adverse Reactions ,Chemistry ,Drug discovery ,Mass spectrometry ,Mass spectrometry imaging ,Mass Spectrometry ,Analytical Chemistry ,Drug development ,Pharmacokinetics ,Pharmaceutical Preparations ,Drug Discovery ,Toxicity Tests ,Animals ,Humans - Published
- 2014
29. Direct targeted quantitative molecular imaging of neurotransmitters in brain tissue sections
- Author
-
Per Svenningsson, Nicoletta Schintu, Richard J. A. Goodwin, Anna Nilsson, Mohammadreza Shariatgorji, Per E. Andrén, Patrik Källback, Alan R. Crossman, Erwan Bezard, Xiaoqun Zhang, Department of Pharmaceutical Biosciences, Biomolecular Imaging and Proteomics, National Center for Mass Spectrometry Imaging, Uppsala University, AstraZeneca, Department of Neurology and Clinical Neuroscience, Center for Molecular Medicine, Karolinska Institutet and Karolinska University Hospital, Motac Neuroscience, Institut des Maladies Neurodégénératives [Bordeaux] (IMN), and Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Neurotransmitter Agents ,Chemistry ,General Neuroscience ,Neuroscience(all) ,[SDV]Life Sciences [q-bio] ,Brain ,Brain tissue ,Mass spectrometry ,Molecular Imaging ,Rats ,Matrix-assisted laser desorption/ionization ,Mice ,Neuroimaging ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Animals ,Molecular imaging ,Neuroscience - Abstract
International audience; Current neuroimaging techniques have very limited abilities to directly identify and quantify neurotransmitters from brain sections. We have developed a molecular-specific approach for the simultaneous imaging and quantitation of multiple neurotransmitters, precursors, and metabolites, such as tyrosine, tryptamine, tyramine, phenethylamine, dopamine, 3-methoxytyramine, serotonin, GABA, glutamate, acetylcholine, and L-alpha-glycerylphosphorylcholine, in histological tissue sections at high spatial resolutions. The method is employed to directly measure changes in the absolute and relative levels of neurotransmitters in specific brain structures in animal disease models and in response to drug treatments, demonstrating the power of mass spectrometry imaging in neuroscience.
- Published
- 2014
30. Mass spectrometry imaging of cassette-dosed drugs for higher throughput pharmacokinetic and biodistribution analysis
- Author
-
James W. Tucker, C. Logan Mackay, Zoltan Takats, Per E. Andrén, Diego Cobice, Anna Nilsson, Nicole Strittmatter, Peter J. H. Webborn, Richard J. A. Goodwin, John G. Swales, and Malcolm R. Clench
- Subjects
Male ,Biodistribution ,TISSUE-SECTIONS ,Administration, Oral ,Pharmacology ,Tandem mass spectrometry ,Mass spectrometry ,METABOLITES ,Mass spectrometry imaging ,MALDI-MS ,Analytical Chemistry ,chemistry.chemical_compound ,Mice ,Pharmacokinetics ,Oral administration ,Tandem Mass Spectrometry ,Drug Discovery ,0399 Other Chemical Sciences ,Animals ,Tissue Distribution ,Dosing ,Rats, Wistar ,Chromatography ,Science & Technology ,LESA ,Bufuralol ,Chemistry, Analytical ,Chemistry ,chemistry ,Pharmaceutical Preparations ,AUTORADIOGRAPHY ,DISCOVERY ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Physical Sciences ,Administration, Intravenous ,0301 Analytical Chemistry - Abstract
Cassette dosing of compounds for preclinical drug plasma pharmacokinetic analysis has been shown to be a powerful strategy within the pharmaceutical industry for increasing throughput while decreasing the number of animals used. Presented here for the first time is data on the application of a cassette dosing strategy for label-free tissue distribution studies. The aim of the study was to image the spatial distribution of eight nonproprietary drugs (haloperidol, bufuralol, midazolam, clozapine, terfenadine, erlotinib, olanzapine, and moxifloxacin) in multiple tissues after oral and intravenous cassette dosing (four compounds per dose route). An array of mass spectrometry imaging technologies, including matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI), liquid extraction surface analysis tandem mass spectrometry (LESA-MS/MS), and desorption electrospray ionization mass spectrometry (DESI-MS) was used. Tissue analysis following intravenous and oral administration of discretely and cassette-dosed compounds demonstrated similar relative abundances across a range of tissues indicating that a cassette dosing approach was applicable. MALDI MSI was unsuccessful in detecting all of the target compounds; therefore, DESI MSI, a complementary mass spectrometry imaging technique, was used to detect additional target compounds. In addition, by adapting technology used for tissue profiling (LESA-MS/MS) low spatial resolution mass spectrometry imaging (∼1 mm) was possible for all targets across all tissues. This study exemplifies the power of multiplatform MSI analysis within a pharmaceutical research and development (R&D) environment. Furthermore, we have illustrated that the cassette dosing approach can be readily applied to provide combined, label-free pharmacokinetic and drug distribution data at an early stage of the drug discovery/development process while minimizing animal usage.
- Published
- 2014
31. Mass Spectrometry Imaging for Dissecting Steroid Intracrinology within Target Tissues
- Author
-
C. Logan Mackay, Ruth Andrew, Diego Cobice, Brian R. Walker, Patrick R. R. Langridge-Smith, Andrew McBride, Richard J. A. Goodwin, and Scott P. Webster
- Subjects
Male ,Resolution (mass spectrometry) ,Collision-induced dissociation ,medicine.medical_treatment ,Tandem mass spectrometry ,Mass spectrometry ,Mass spectrometry imaging ,Fourier transform ion cyclotron resonance ,Article ,Mass Spectrometry ,Analytical Chemistry ,Steroid ,Rats, Sprague-Dawley ,Mice ,Tandem Mass Spectrometry ,Adrenal Glands ,medicine ,Animals ,Brain Chemistry ,Mice, Knockout ,Chromatography ,Chemistry ,Brain ,Rats ,Mice, Inbred C57BL ,Matrix-assisted laser desorption/ionization ,Biochemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Steroids ,Chromatography, Liquid - Abstract
Steroid concentrations within tissues are modulated by intracellular enzymes. Such ‘steroid intracrinology’ influences hormone-dependent cancers and obesity, and provides targets for pharmacological inhibition. However, no high resolution methods exist to quantify steroids within target tissues. We developed mass spectrometry imaging (MSI), combining matrix assisted laser desorption ionization with on-tissue derivatization with Girard T and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry, to quantify substrate and product (11-dehydrocorticosterone and corticosterone) of the glucocorticoid-amplifying enzyme 11β-HSD1. Regional steroid distribution was imaged at 150-200μm resolution in rat adrenal gland and mouse brain sections, and confirmed with collision induced dissociation/liquid extraction surface analysis. In brains of mice with 11β-HSD1 deficiency or inhibition, MSI quantified changes in sub-regional corticosterone/11-dehydrocorticosterone ratio, distribution of inhibitor, and accumulation of the alternative 11β-HSD1 substrate, 7-ketocholesterol. MSI data correlated well with LC-MS/MS in whole brain homogenates. MSI with derivatization is a powerful new tool to investigate steroid biology within tissues.
- Published
- 2013
32. Sample preparation for mass spectrometry imaging: small mistakes can lead to big consequences
- Author
-
Richard J. A. Goodwin
- Subjects
Diagnostic Imaging ,Standardization ,Medical Errors ,business.industry ,Chemistry ,Biophysics ,Nanotechnology ,Microtomy ,Mass spectrometry ,Biochemistry ,Mass spectrometry imaging ,Mass Spectrometry ,Specimen Handling ,Research Design ,Data quality ,Animals ,Humans ,Sample preparation ,Sample collection ,Instrumentation (computer programming) ,Process engineering ,business ,Algorithms - Abstract
Mass spectrometry imaging (MSI) enables the direct analysis of molecules from the surface of a wide variety of samples, allowing the multiplex measurement of both abundance and distribution of small molecules, lipids, peptides and proteins. As the technology has been refined an increasing number of ionization methods and mass analyzers has been used that enable increased spatial and spectral resolution measurements to be made at an increased speed. Alongside the instrumentation improvements there has been optimization of sample preparation procedures that allow the highest quality data to be obtained, reproducibly, from an ever increasing diversity of samples. This review will consider the development and standardization of sample preparation methods applicable to MSI, describing the stages and procedures undertaken from the instance of sample collection, through storage, preparation and on through final processing prior to analysis. Recent technical advancements will be highlighted and areas where further experimentation and optimization may well be required will be described. All aspects of the sample preparation pipeline will be considered in detail, with examples from the literature used to emphasize why rigorous sample preparation for MSI is vital to achieve the most accurate, reproducible and validated MSI data possible.
- Published
- 2012
33. In Situ Mass Spectrometry Imaging and Ex Vivo Characterization of Renal Crystalline Deposits Induced in Multiple Preclinical Drug Toxicology Studies
- Author
-
Alexander Svanhagen, Johan Lindberg, Dennis Hellgren, Per E. Andrén, Richard J. A. Goodwin, Sivert Bjurström, Elisa Basmaci, Benita Forngren, Anna Nilsson, Anita Annas, and Ingela Gustafsson
- Subjects
Drugs and Devices ,Medicin och hälsovetenskap ,Magnetic Resonance Spectroscopy ,Drug Research and Development ,Metabolite ,Toxic Agents ,lcsh:Medicine ,Pharmacology ,Mass spectrometry ,Kidney ,Toxicology ,Biochemistry ,Medical and Health Sciences ,Mass spectrometry imaging ,Mass Spectrometry ,chemistry.chemical_compound ,Model Organisms ,Liquid chromatography–mass spectrometry ,Drug Discovery ,Animals ,Enzyme Inhibitors ,lcsh:Science ,Biology ,Prostaglandin-E Synthases ,Multidisciplinary ,Chromatography ,Chemistry ,lcsh:R ,Clinical Pharmacology ,Kidney metabolism ,Animal Models ,Rats ,Intramolecular Oxidoreductases ,Drug development ,Small Molecules ,Rat ,Medicine ,lcsh:Q ,Female ,Ex vivo ,Drug metabolism ,Chromatography, Liquid ,Research Article - Abstract
Drug toxicity observed in animal studies during drug development accounts for the discontinuation of many drug candidates, with the kidney being a major site of tissue damage. Extensive investigations are often required to reveal the mechanisms underlying such toxicological events and in the case of crystalline deposits the chemical composition can be problematic to determine. In the present study, we have used mass spectrometry imaging combined with a set of advanced analytical techniques to characterize such crystalline deposits in situ. Two potential microsomal prostaglandin E synthase 1 inhibitors, with similar chemical structure, were administered to rats over a seven day period. This resulted in kidney damage with marked tubular degeneration/regeneration and crystal deposits within the tissue that was detected by histopathology. Results from direct tissue section analysis by matrix-assisted laser desorption ionization mass spectrometry imaging were combined with data obtained following manual crystal dissection analyzed by liquid chromatography mass spectrometry and nuclear magnetic resonance spectroscopy. The chemical composition of the crystal deposits was successfully identified as a common metabolite, bisulphonamide, of the two drug candidates. In addition, an un-targeted analysis revealed molecular changes in the kidney that were specifically associated with the area of the tissue defined as pathologically damaged. In the presented study, we show the usefulness of combining mass spectrometry imaging with an array of powerful analytical tools to solve complex toxicological problems occurring during drug development. De två första författarna delar förstaförfattarskapet.De två sista författarna delar sistaförfattarskapet.
- Published
- 2012
34. Mass spectrometry imaging of pharmacological compounds in tissue sections
- Author
-
Andrew R. Pitt and Richard J. A. Goodwin
- Subjects
Analyte ,Histocytological Preparation Techniques ,Resolution (mass spectrometry) ,Chemistry ,Clinical Biochemistry ,General Medicine ,Computational biology ,Mass spectrometry ,Bioinformatics ,Mass spectrometry imaging ,Mass Spectrometry ,Analytical Chemistry ,Molecular Imaging ,Medical Laboratory Technology ,Tissue sections ,Pharmaceutical Preparations ,Animals ,Humans ,Sample preparation ,General Pharmacology, Toxicology and Pharmaceutics ,Molecular imaging - Abstract
The use of MS imaging (MSI) to resolve the spatial and pharmacodynamic distributions of compounds in tissues is emerging as a powerful tool for pharmacological research. Unlike established imaging techniques, only limited a priori knowledge is required and no extensive manipulation (e.g., radiolabeling) of drugs is necessary prior to dosing. MS provides highly multiplexed detection, making it possible to identify compounds, their metabolites and other changes in biomolecular abundances directly off tissue sections in a single pass. This can be employed to obtain near cellular, or potentially subcellular, resolution images. Consideration of technical limitations that affect the process is required, from sample preparation through to analyte ionization and detection. The techniques have only recently been adapted for imaging and novel variations to the established MSI methodologies will further enhance the application of MSI for pharmacological research.
- Published
- 2010
35. Protein and peptides in pictures: imaging with MALDI mass spectrometry
- Author
-
Stephen R. Pennington, Andrew R. Pitt, and Richard J. A. Goodwin
- Subjects
Chemistry ,Spatially resolved ,Disease progression ,Proteins ,Computational biology ,Proteomics ,Mass spectrometry ,Biochemistry ,Peptide Mapping ,Mass spectrometry imaging ,Matrix-assisted laser desorption/ionization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Animals ,Humans ,Identification (biology) ,Tissue Preservation ,Biomarker discovery ,Peptides ,Molecular Biology ,Biomarkers - Abstract
Imaging using MS has the potential to deliver highly parallel, multiplexed data on the specific localization of molecular ions in tissue samples directly, and to measure and map the variations of these ions during development and disease progression or treatment. There is an intrinsic potential to be able to identify the biomarkers in the same experiment, or by relatively simple extension of the technique. Unlike many other imaging techniques, no a priori knowledge of the markers being sought is necessary. This review concentrates on the use of MALDI-MS for MS imaging (MSI) of proteins and peptides, with an emphasis on mammalian tissue. We discuss the methodologies used, their potential limitations, overall experimental considerations and progress that has been made towards establishing MALDI-MSI as a routine technique for the spatially resolved measurement of peptides and proteins. As well as determining the local abundance of individual molecular ions, there is the potential to determine their identity within the same experiment using relatively simple extensions of the basic techniques. In this way MSI offers an important opportunity for biomarker discovery and identification.
- Published
- 2008
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