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Matrix assisted laser desorption ionisation mass spectrometry imaging (MALDI-MSI), was used to obtain images of lipids and metabolite distribution in formalin fixed and embedded in paraffin (FFPE) whole eye sections containing primary uveal melanomas (UM). Using this technique, it was possible to obtain images of lysophosphatidylcholine (LPC) type lipid distribution that highlighted the tumour regions. Laser ablation inductively coupled plasma mass spectrometry images (LA-ICP-MS) performed on UM sections showed increases in copper within the tumour periphery and intratumoural zinc in tissue from patients with poor prognosis. These preliminary data indicate that multi-modal MSI has the potential to provide insights into the role of trace metals and cancer metastasis.
Laura M. Cole; Joshua Handley; Emmanuelle Claude; Catherine J. Duckett; Hardeep S. Mudhar; Karen Sisley; Malcolm R. Clench. Multi-Modal Mass Spectrometric Imaging of Uveal Melanoma. Metabolites 2021, 11, 560 .
AMA StyleLaura M. Cole, Joshua Handley, Emmanuelle Claude, Catherine J. Duckett, Hardeep S. Mudhar, Karen Sisley, Malcolm R. Clench. Multi-Modal Mass Spectrometric Imaging of Uveal Melanoma. Metabolites. 2021; 11 (8):560.
Chicago/Turabian StyleLaura M. Cole; Joshua Handley; Emmanuelle Claude; Catherine J. Duckett; Hardeep S. Mudhar; Karen Sisley; Malcolm R. Clench. 2021. "Multi-Modal Mass Spectrometric Imaging of Uveal Melanoma." Metabolites 11, no. 8: 560.
Osteosarcoma (OS) is the most common primary bone malignancy and largely effects adolescents and young adults, with 60% of patients under the age of 25. There are multiple cell models of OS described in vitro that express the specific genetic alterations of the sarcoma. In the work reported here, multiple mass spectrometry imaging (MSI) modalities were employed to characterise two aggregated cellular models of OS models formed using the MG63 and SAOS-2 cell lines. Phenotyping of the metabolite activity within the two OS aggregoid models was achieved and a comparison of the metabolite data with OS human tissue samples revealed relevant fatty acid and phospholipid markers. Although, annotations of these species require MS/MS analysis for confident identification of the metabolites. From the putative assignments however, it was suggested that the MG63 aggregoids are an aggressive tumour model that exhibited metastatic-like potential. Alternatively, the SAOS-2 aggregoids are more mature osteoblast-like phenotype that expressed characteristics of cellular differentiation and bone development. It was determined the two OS aggregoid models shared similarities of metabolic behaviour with different regions of OS human tissues, specifically of the higher metastatic grade.
Lucy Flint; Gregory Hamm; Joseph Ready; Stephanie Ling; Catherine Duckett; Neil Cross; Laura Cole; David Smith; Richard Goodwin; Malcolm Clench. Comparison of Osteosarcoma Aggregated Tumour Models with Human Tissue by Multimodal Mass Spectrometry Imaging. Metabolites 2021, 11, 506 .
AMA StyleLucy Flint, Gregory Hamm, Joseph Ready, Stephanie Ling, Catherine Duckett, Neil Cross, Laura Cole, David Smith, Richard Goodwin, Malcolm Clench. Comparison of Osteosarcoma Aggregated Tumour Models with Human Tissue by Multimodal Mass Spectrometry Imaging. Metabolites. 2021; 11 (8):506.
Chicago/Turabian StyleLucy Flint; Gregory Hamm; Joseph Ready; Stephanie Ling; Catherine Duckett; Neil Cross; Laura Cole; David Smith; Richard Goodwin; Malcolm Clench. 2021. "Comparison of Osteosarcoma Aggregated Tumour Models with Human Tissue by Multimodal Mass Spectrometry Imaging." Metabolites 11, no. 8: 506.
Despite being a critical molecule in the brain, mass spectrometry imaging (MSI) of cholesterol has been under-reported compared to other lipids due to the difficulty in ionizing the sterol molecule. In the present work, we have employed an on-tissue enzyme-assisted derivatization strategy to improve detection of cholesterol in brain tissue sections. We report distribution and levels of cholesterol across specific structures of the mouse brain, in a model of Niemann-Pick type C1 disease, and during brain development. MSI revealed that in the adult mouse, cholesterol is the highest in the pons and medulla and how its distribution changes during development. Cholesterol was significantly reduced in the corpus callosum and other brain regions in the Npc1 null mouse, confirming hypomyelination at the molecular level. Our study demonstrates the potential of MSI to the study of sterols in neuroscience.
Roberto Angelini; Eylan Yutuc; Mark F. Wyatt; Jillian Newton; Fowzi A. Yusuf; Lauren Griffiths; Benjamin J. Cooze; Dana El Assad; Gilles Frache; Wei Rao; Luke B. Allen; Zeljka Korade; Thu T. A. Nguyen; Rathnayake A. C. Rathnayake; Stephanie M. Cologna; Owain W. Howell; Malcolm R. Clench; Yuqin Wang; William J. Griffiths. Visualizing Cholesterol in the Brain by On-Tissue Derivatization and Quantitative Mass Spectrometry Imaging. Analytical Chemistry 2021, 93, 4932 -4943.
AMA StyleRoberto Angelini, Eylan Yutuc, Mark F. Wyatt, Jillian Newton, Fowzi A. Yusuf, Lauren Griffiths, Benjamin J. Cooze, Dana El Assad, Gilles Frache, Wei Rao, Luke B. Allen, Zeljka Korade, Thu T. A. Nguyen, Rathnayake A. C. Rathnayake, Stephanie M. Cologna, Owain W. Howell, Malcolm R. Clench, Yuqin Wang, William J. Griffiths. Visualizing Cholesterol in the Brain by On-Tissue Derivatization and Quantitative Mass Spectrometry Imaging. Analytical Chemistry. 2021; 93 (11):4932-4943.
Chicago/Turabian StyleRoberto Angelini; Eylan Yutuc; Mark F. Wyatt; Jillian Newton; Fowzi A. Yusuf; Lauren Griffiths; Benjamin J. Cooze; Dana El Assad; Gilles Frache; Wei Rao; Luke B. Allen; Zeljka Korade; Thu T. A. Nguyen; Rathnayake A. C. Rathnayake; Stephanie M. Cologna; Owain W. Howell; Malcolm R. Clench; Yuqin Wang; William J. Griffiths. 2021. "Visualizing Cholesterol in the Brain by On-Tissue Derivatization and Quantitative Mass Spectrometry Imaging." Analytical Chemistry 93, no. 11: 4932-4943.
Introduction Three-dimensional (3D) cell cultures have become increasingly important materials to investigate biological processes and drug efficacy and toxicity. The ability of 3D cultures to mimic the physiology of primary tissues and organs in the human body enables further insight into cellular behaviour and is hence highly desirable in early stage drug development. Analysing the spatial distribution of drug compounds and endogenous molecules provides an insight into the efficacy of a drug whilst simultaneously giving information on biological responses. Areas Covered In this review we will examine the main 3D cell culture systems employed and applications, which describe their integration with mass spectrometry imaging (MSI). Expert Opinion MSI is a powerful technique that can map a vast range of molecules simultaneously in tissues without the addition of labels that can provide insights into the efficacy and safety of a new drug. The combination of MSI and 3D cell cultures has emerged as a promising tool in early stage drug analysis. However, the most common administration route for pharmaceutical drugs is via oral delivery. The use of MSI in combination with models of the GI tract is an area that has been little explored to date, the reasons for this are discussed.
Chloe E Spencer; Lucy E Flint; Catherine J Duckett; Laura M Cole; Neil Cross; David P Smith; Malcolm R Clench. Role of MALDI-MSI in combination with 3D tissue models for early stage efficacy and safety testing of drugs and toxicants. Expert Review of Proteomics 2020, 17, 827 -841.
AMA StyleChloe E Spencer, Lucy E Flint, Catherine J Duckett, Laura M Cole, Neil Cross, David P Smith, Malcolm R Clench. Role of MALDI-MSI in combination with 3D tissue models for early stage efficacy and safety testing of drugs and toxicants. Expert Review of Proteomics. 2020; 17 (11-12):827-841.
Chicago/Turabian StyleChloe E Spencer; Lucy E Flint; Catherine J Duckett; Laura M Cole; Neil Cross; David P Smith; Malcolm R Clench. 2020. "Role of MALDI-MSI in combination with 3D tissue models for early stage efficacy and safety testing of drugs and toxicants." Expert Review of Proteomics 17, no. 11-12: 827-841.
SummaryDespite being a critical molecule for neurobiology and brain health, mass spectrometry imaging (MSI) of cholesterol has been under reported compared to other lipids, due to the difficulty in ionising the sterol molecule. In the present work we have employed an on-tissue enzyme-assisted derivatisation strategy to improve detection of cholesterol in brain tissue sections. We report distribution and levels of cholesterol across specific brain structures of the mouse brain, in a model of Niemann-Pick type C1 (NPC1) disease, and during brain development. MSI revealed how cholesterol changes during development and that in the adult is highest in pons and medulla of the brain stem. Cholesterol was significantly reduced in the corpus callosum and other brain regions in the Npc1 null mouse, confirming hypomyelination at the molecular level. Our study demonstrates the potential of MSI to the study of sterols in neuroscience.
Roberto Angelini; Eylan Yutuc; Mark F Wyatt; Jillian Newton; Fowzi Adam Yusuf; Lauren Griffiths; Benjamin Jordan Cooze; Dana El Assad; Gilles Frache; Wei Rao; Luke B. Allen; Zeljka Korade; Thu Ta Nguyen; Rathnayake Ac Rathnayake; Stephanie M Cologna; Owain W Howell; Malcolm R Clench; Yuqin Wang; William J Griffiths. Visualising Cholesterol in Brain by On-Tissue Derivatisation and Quantitative Mass Spectrometry Imaging. 2020, 1 .
AMA StyleRoberto Angelini, Eylan Yutuc, Mark F Wyatt, Jillian Newton, Fowzi Adam Yusuf, Lauren Griffiths, Benjamin Jordan Cooze, Dana El Assad, Gilles Frache, Wei Rao, Luke B. Allen, Zeljka Korade, Thu Ta Nguyen, Rathnayake Ac Rathnayake, Stephanie M Cologna, Owain W Howell, Malcolm R Clench, Yuqin Wang, William J Griffiths. Visualising Cholesterol in Brain by On-Tissue Derivatisation and Quantitative Mass Spectrometry Imaging. . 2020; ():1.
Chicago/Turabian StyleRoberto Angelini; Eylan Yutuc; Mark F Wyatt; Jillian Newton; Fowzi Adam Yusuf; Lauren Griffiths; Benjamin Jordan Cooze; Dana El Assad; Gilles Frache; Wei Rao; Luke B. Allen; Zeljka Korade; Thu Ta Nguyen; Rathnayake Ac Rathnayake; Stephanie M Cologna; Owain W Howell; Malcolm R Clench; Yuqin Wang; William J Griffiths. 2020. "Visualising Cholesterol in Brain by On-Tissue Derivatisation and Quantitative Mass Spectrometry Imaging." , no. : 1.
Mass spectrometry imaging (MSI) is a powerful and versatile technique able to investigate the spatial distribution of multiple non-labelled endogenous and exogenous analytes simultaneously, within a wide range of samples. Over the last two decades, MSI has found widespread application for an extensive range of disciplines including pre-clinical drug discovery, clinical applications and human identification for forensic purposes. Technical advances in both instrumentation and software capabilities have led to a continual increase in the interest in MSI; however, there are still some limitations. In this review, we discuss the emerging applications in MSI that significantly impact three key areas of mass spectrometry (MS) research—clinical, pre-clinical and forensics—and roadblocks to the expansion of use of MSI in these areas.
Cristina Russo; Cameron Heaton; Lucy Flint; Oana Voloaca; Sarah Haywood-Small; Malcolm Ronald Clench; Simona Francese; Laura Margaret Cole. Emerging applications in mass spectrometry imaging; enablers and roadblocks. Journal of Spectral Imaging 2020, 9, 1 .
AMA StyleCristina Russo, Cameron Heaton, Lucy Flint, Oana Voloaca, Sarah Haywood-Small, Malcolm Ronald Clench, Simona Francese, Laura Margaret Cole. Emerging applications in mass spectrometry imaging; enablers and roadblocks. Journal of Spectral Imaging. 2020; 9 ():1.
Chicago/Turabian StyleCristina Russo; Cameron Heaton; Lucy Flint; Oana Voloaca; Sarah Haywood-Small; Malcolm Ronald Clench; Simona Francese; Laura Margaret Cole. 2020. "Emerging applications in mass spectrometry imaging; enablers and roadblocks." Journal of Spectral Imaging 9, no. : 1.
Rationale Matrix‐assisted laser desorption/ionization‐mass spectrometry imaging (MALDI‐MSI) is routinely employed to monitor the distribution of compounds in tissue sections and generate 2D images. Whilst informative the images do not represent the distribution of the analyte of interest through the entire organ. The generation of 3D images is an exciting field that can provide a deeper view of the analyte of interest throughout an entire organ. Methods Serial sections of mouse and rat lung tissue were obtained at 120 μm depth intervals and imaged individually. Homogenate registration markers were incorporated in order to aid the final 3D image construction. Using freely available software packages, the images were stacked together to generate a 3D image that showed the distribution of endogenous species throughout the lungs. Results Preliminary tests were performed on 16 serial tissue sections of mouse lungs. A 3D model showing the distribution of phosphocholine at m/z 184.09 was constructed, which defined the external structure of the lungs and trachea. Later, a second experiment was performed using 24 serial tissue sections of the left lung of a rat. Two molecular markers, identified as [PC (32:1)+K]+ at m/z 770.51 and [PC (36:4)+K]+ at m/z 820.52 were used to generate 3D models of the parenchyma and airways, respectively. Conclusions A straightforward method to generate 3D MALDI‐MS images of selected molecules in lung tissue has been presented. Using freely available imaging software, the 3D distributions of molecules related to different anatomical features were determined.
Bryn Flinders; Josie Morrell; Peter S. Marshall; Lisa E. Ranshaw; Ron M.A. Heeren; Malcolm R. Clench. Monitoring the three‐dimensional distribution of endogenous species in the lungs by matrix‐assisted laser desorption/ionization mass spectrometry imaging. Rapid Communications in Mass Spectrometry 2020, 35, 1 .
AMA StyleBryn Flinders, Josie Morrell, Peter S. Marshall, Lisa E. Ranshaw, Ron M.A. Heeren, Malcolm R. Clench. Monitoring the three‐dimensional distribution of endogenous species in the lungs by matrix‐assisted laser desorption/ionization mass spectrometry imaging. Rapid Communications in Mass Spectrometry. 2020; 35 (1):1.
Chicago/Turabian StyleBryn Flinders; Josie Morrell; Peter S. Marshall; Lisa E. Ranshaw; Ron M.A. Heeren; Malcolm R. Clench. 2020. "Monitoring the three‐dimensional distribution of endogenous species in the lungs by matrix‐assisted laser desorption/ionization mass spectrometry imaging." Rapid Communications in Mass Spectrometry 35, no. 1: 1.
Mass spectrometry imaging (MSI) is an established analytical tool capable of defining and understanding complex tissues by determining the spatial distribution of biological molecules. Three-dimensional (3D) cell culture models mimic the pathophysiological environment of in vivo tumors and are rapidly emerging as a valuable research tool. Here, multimodal MSI techniques were employed to characterize a novel aggregated 3D lung adenocarcinoma model, developed by the group to mimic the in vivo tissue. Regions of tumor heterogeneity and the hypoxic microenvironment were observed based on the spatial distribution of a variety of endogenous molecules. Desorption electrospray ionization (DESI)-MSI defined regions of a hypoxic core and a proliferative outer layer from metabolite distribution. Targeted metabolites (e.g. lactate, glutamine and citrate) were mapped to pathways of glycolysis and the TCA cycle demonstrating tumor metabolic behavior. The first application of imaging mass cytometry (IMC) with 3D cell culture enabled single-cell phenotyping at 1 µm spatial resolution. Protein markers of proliferation (Ki-67) and hypoxia (glucose transporter 1) defined metabolic signaling in the aggregoid model, which complemented the metabolite data. Laser ablation inductively coupled plasma (LA-ICP)-MSI analysis localized endogenous elements including magnesium and copper, further differentiating the hypoxia gradient and validating the protein expression. Obtaining a large amount of molecular information of a complementary nature enabled an in-depth understanding of the biological processes within the novel tumor model. Combining powerful imaging techniques to characterize the aggregated 3D culture highlighted a future methodology with potential applications in cancer research and drug development.
Lucy Ellen Flint; Gregory R. Hamm; Joseph Ready; Stephanie Ling; Catherine Jane Duckett; Neil Alan Cross; Laura Margaret Cole; David P. Smith; Richard J. A. Goodwin; Malcolm R. Clench. Characterization of an Aggregated Three-Dimensional Cell Culture Model by Multimodal Mass Spectrometry Imaging. Analytical Chemistry 2020, 92, 12538 -12547.
AMA StyleLucy Ellen Flint, Gregory R. Hamm, Joseph Ready, Stephanie Ling, Catherine Jane Duckett, Neil Alan Cross, Laura Margaret Cole, David P. Smith, Richard J. A. Goodwin, Malcolm R. Clench. Characterization of an Aggregated Three-Dimensional Cell Culture Model by Multimodal Mass Spectrometry Imaging. Analytical Chemistry. 2020; 92 (18):12538-12547.
Chicago/Turabian StyleLucy Ellen Flint; Gregory R. Hamm; Joseph Ready; Stephanie Ling; Catherine Jane Duckett; Neil Alan Cross; Laura Margaret Cole; David P. Smith; Richard J. A. Goodwin; Malcolm R. Clench. 2020. "Characterization of an Aggregated Three-Dimensional Cell Culture Model by Multimodal Mass Spectrometry Imaging." Analytical Chemistry 92, no. 18: 12538-12547.
RATIONALE Malignant pleural mesothelioma is an extremely aggressive and incurable malignancy associated with prior exposure to asbestos fibres. Difficulties remain in relation to early diagnosis, notably due to impeded identification of asbestos in lung tissue. This study describes a novel laser ablation inductively coupled plasma mass spectrometry (LA‐ICP‐MS) imaging approach to identify asbestos within mesothelioma models with clinical significance. METHODS Human mesothelioma cells were exposed to different types of asbestos fibres and prepared on plastic slides for LA‐ICP‐MS analysis. No further sample preparation was required prior to analysis, which was performed using an NWR Image 266nm laser ablation system coupled to an Element XR sector‐field ICP mass spectrometer, with a lateral resolution of 2 μm. Data was processed using LA‐ICP‐MS ImageTool v1.7 with the final graphic production made using DPlot Software. RESULTS Four different mineral fibres were successfully identified within the mesothelioma samples based on some of the most abundant elements that make up these fibres (Si, Mg and Fe). Using LA‐ICP‐MS as an imaging tool provided information on the spatial distribution of the fibres at cellular level, which is essential in asbestos detection within tissue samples. Based on the metal counts generated by the different types of asbestos, different fibres can be identified based on shape, size, and elemental composition. Detection of Ca was attempted but requires further optimisation. CONCLUSION Asbestos fibres detection in the lung tissues is very useful, if not necessary, to complete the pathological diagnosis of asbestos‐related malignancies in medicolegal field. For the first time, this study demonstrates the successful application of LA‐ICP‐MS imaging to identify asbestos fibres and other mineral fibres within mesothelioma samples. Ultimately, high‐resolution, fast‐speed LA‐ICP‐MS analysis has the potential to be integrated into clinical workflow to aid earlier detection and stratification of mesothelioma patient samples.
Oana M. Voloaca; Calum J. Greenhalgh; Laura M. Cole; Malcolm R. Clench; Amy J. Managh; Sarah L. Haywood‐Small. Laser ablation inductively coupled plasma mass spectrometry as a novel clinical imaging tool to detect asbestos fibres in malignant mesothelioma. Rapid Communications in Mass Spectrometry 2020, 34, 1 .
AMA StyleOana M. Voloaca, Calum J. Greenhalgh, Laura M. Cole, Malcolm R. Clench, Amy J. Managh, Sarah L. Haywood‐Small. Laser ablation inductively coupled plasma mass spectrometry as a novel clinical imaging tool to detect asbestos fibres in malignant mesothelioma. Rapid Communications in Mass Spectrometry. 2020; 34 (21):1.
Chicago/Turabian StyleOana M. Voloaca; Calum J. Greenhalgh; Laura M. Cole; Malcolm R. Clench; Amy J. Managh; Sarah L. Haywood‐Small. 2020. "Laser ablation inductively coupled plasma mass spectrometry as a novel clinical imaging tool to detect asbestos fibres in malignant mesothelioma." Rapid Communications in Mass Spectrometry 34, no. 21: 1.
Dysregulated cholesterol metabolism is implicated in a number of neurological disorders. Many sterols, including cholesterol and its precursors and metabolites, are biologically active and important for proper brain function. However, spatial cholesterol metabolism in brain and the resulting sterol distributions are poorly defined. To better understand cholesterol metabolism in situ across the complex functional regions of brain, we have developed on-tissue enzyme-assisted derivatization in combination with microliquid extraction for surface analysis and liquid chromatography-mass spectrometry to locate sterols in tissue slices (10 µm) of mouse brain. The method provides sterolomic analysis at 400-µm spot diameter with a limit of quantification of 0.01 ng/mm2. It overcomes the limitations of previous mass spectrometry imaging techniques in analysis of low-abundance and difficult-to-ionize sterol molecules, allowing isomer differentiation and structure identification. Here we demonstrate the spatial distribution and quantification of multiple sterols involved in cholesterol metabolic pathways in wild-type andcholesterol 24S-hydroxylaseknockout mouse brain. The technology described provides a powerful tool for future studies of spatial cholesterol metabolism in healthy and diseased tissues.
Eylan Yutuc; Roberto Angelini; Mark Baumert; Natalia Mast; Irina Pikuleva; Jillian Newton; Malcolm R. Clench; David O. F. Skibinski; Owain W. Howell; Yuqin Wang; William J. Griffiths. Localization of sterols and oxysterols in mouse brain reveals distinct spatial cholesterol metabolism. Proceedings of the National Academy of Sciences 2020, 117, 5749 -5760.
AMA StyleEylan Yutuc, Roberto Angelini, Mark Baumert, Natalia Mast, Irina Pikuleva, Jillian Newton, Malcolm R. Clench, David O. F. Skibinski, Owain W. Howell, Yuqin Wang, William J. Griffiths. Localization of sterols and oxysterols in mouse brain reveals distinct spatial cholesterol metabolism. Proceedings of the National Academy of Sciences. 2020; 117 (11):5749-5760.
Chicago/Turabian StyleEylan Yutuc; Roberto Angelini; Mark Baumert; Natalia Mast; Irina Pikuleva; Jillian Newton; Malcolm R. Clench; David O. F. Skibinski; Owain W. Howell; Yuqin Wang; William J. Griffiths. 2020. "Localization of sterols and oxysterols in mouse brain reveals distinct spatial cholesterol metabolism." Proceedings of the National Academy of Sciences 117, no. 11: 5749-5760.
Matrix-Assisted Ionisation in Vacuum (MAIV) is a new ionisation technique which ionises non-volatile compounds producing electrospray ionisation-like spectra. Its simple, matrix-assisted laser desorption/ionisation-like sample preparation allows for rapid analysis, with no requirement for external energy in the form of a laser or high voltage to produce ions. Ionisation occurs when the matrix (often 3-nitrobenzonitrile) is exposed to sub-ambient pressure. Here, the first use of this revolutionary new ionisation technique to image biological samples is reported. A commercial quadrupole-quadrupole-time-of-flight mass spectrometer was modified to incorporate control of the ion source pressure and a reduced sampling cone orifice diameter. In initial experiments, optimisation of source pressure and matrix composition was carried out to increase the longevity of ion formation. It was noted during these experiments that ion production was only observed when the sample was directly under the sampling cone. Optimisation of sample extraction into the MAIV matrix by the addition of 5% chloroform enabled MAIV mass spectrometry imaging of lipids in rat brain sections to be carried out in raster imaging mode. Modification of the size and position of the sampling cone improved the selectivity obtainable in these images. Although the quality of these initial images is relatively poor, work is underway to improve the spatial resolution by further modification of the ion source and progress is reported.
Alex Harding; James Hough; Charlotte Curtis; Daniel Kinsman; Malcolm Clench. Matrix-assisted ionisation in vacuum mass spectrometry and imaging on a modified quadrupole-quadrupole-time-of-flight mass spectrometer. Journal of Spectral Imaging 2019, 1 .
AMA StyleAlex Harding, James Hough, Charlotte Curtis, Daniel Kinsman, Malcolm Clench. Matrix-assisted ionisation in vacuum mass spectrometry and imaging on a modified quadrupole-quadrupole-time-of-flight mass spectrometer. Journal of Spectral Imaging. 2019; ():1.
Chicago/Turabian StyleAlex Harding; James Hough; Charlotte Curtis; Daniel Kinsman; Malcolm Clench. 2019. "Matrix-assisted ionisation in vacuum mass spectrometry and imaging on a modified quadrupole-quadrupole-time-of-flight mass spectrometer." Journal of Spectral Imaging , no. : 1.
Dysregulated cholesterol metabolism is implicated in a number of neurological disorders. Many sterols, including cholesterol and its precursors and metabolites, are biologically active and important for proper brain function. However, spatial cholesterol metabolism in brain and the resulting sterol distributions are poorly defined. To better understand cholesterol metabolism in situ across the complex functional regions of brain, we have developed on-tissue enzyme-assisted derivatisation in combination with micro-liquid-extraction for surface analysis and liquid chromatography - mass spectrometry to image sterols in tissue slices (10 µm) of mouse brain. The method provides sterolomic analysis at 400 µm spot diameter with a limit of quantification of 0.01 ng/mm2. It overcomes the limitations of previous mass spectrometry imaging techniques in analysis of low abundance and difficult to ionise sterol molecules, allowing isomer differentiation and structure identification. Here we demonstrate the spatial distribution and quantification of multiple sterols involved in cholesterol metabolic pathways in wild type and cholesterol 24S-hydroxylase knock-out mouse brain. The technology described provides a powerful tool for future studies of spatial cholesterol metabolism in healthy and diseased tissues.SignificanceThe brain is a remarkably complex organ and cholesterol homeostasis underpins brain function. It is known that cholesterol is not evenly distributed across different brain regions, however, the precise map of cholesterol metabolism in the brain remains unclear. If cholesterol metabolism is to be correlated with brain function it is essential to generate such a map. Here we describe an advanced mass spectrometry imaging platform to reveal spatial cholesterol metabolism in situ at 400 µm resolution on 10 µm tissue slices from mouse brain. We mapped, not only cholesterol, but also other biologically active sterols arising from cholesterol turnover in both wild type and mice lacking cholesterol 24-hydroxylase (Cyp46a1), the major cholesterol metabolising enzyme.
Eylan Yutuc; Roberto Angelini; Mark Baumert; Natalia Mast; Irina Pikuleva; Jillian Newton; Malcolm R. Clench; David O.F. Skibinski; Owain W. Howell; Yuqin Wang; William J. Griffiths. Imaging Sterols and Oxysterols in Mouse Brain Reveals Distinct Spatial Cholesterol Metabolism. 2018, 450973 .
AMA StyleEylan Yutuc, Roberto Angelini, Mark Baumert, Natalia Mast, Irina Pikuleva, Jillian Newton, Malcolm R. Clench, David O.F. Skibinski, Owain W. Howell, Yuqin Wang, William J. Griffiths. Imaging Sterols and Oxysterols in Mouse Brain Reveals Distinct Spatial Cholesterol Metabolism. . 2018; ():450973.
Chicago/Turabian StyleEylan Yutuc; Roberto Angelini; Mark Baumert; Natalia Mast; Irina Pikuleva; Jillian Newton; Malcolm R. Clench; David O.F. Skibinski; Owain W. Howell; Yuqin Wang; William J. Griffiths. 2018. "Imaging Sterols and Oxysterols in Mouse Brain Reveals Distinct Spatial Cholesterol Metabolism." , no. : 450973.
Cristina Russo; Emily E. L. Lewis; Lucy Flint; Malcolm R. Clench. Front Cover: Mass Spectrometry Imaging of 3D Tissue Models. PROTEOMICS 2018, 18, 1 .
AMA StyleCristina Russo, Emily E. L. Lewis, Lucy Flint, Malcolm R. Clench. Front Cover: Mass Spectrometry Imaging of 3D Tissue Models. PROTEOMICS. 2018; 18 (14):1.
Chicago/Turabian StyleCristina Russo; Emily E. L. Lewis; Lucy Flint; Malcolm R. Clench. 2018. "Front Cover: Mass Spectrometry Imaging of 3D Tissue Models." PROTEOMICS 18, no. 14: 1.
The combination of microspotting of analytical and internal standards, matrix sublimation and recently developed software for quantitative mass spectrometry imaging has been used to develop a high resolution method for the determination of Terbinafine hydrochloride in the epidermal region of a full thickness living skin equivalent model. A quantitative assessment of the effect of the addition of the penetration enhancer (dimethyl isosorbide (DMI)) to the delivery vehicle has also been performed and data have been compared to those obtained from LC/MS/MS measurements of homogenates of isolated epidermal tissue. At 10% DMI the levels of signal detected for the drug in the epidermis were 0.20 ± 0.072 mg/g tissue for QMSI and 0.28 ± 0.040 mg/g tissue for LC/MS/MS; at 50% DMI 0.69 ± 0.23 mg/g tissue for QMSI and 0.66 ± 0.057 mg/g tissue for LC/MS/MS. Comparison of means and standard deviations indicates no significant difference between the values obtained by the two methods.
Cristina Russo; Neil Brickelbank; Catherine Duckett; Steve Mellor; Stephen Rumbelow; Malcolm R. Clench; Neil Bricklebank. Quantitative Investigation of Terbinafine Hydrochloride Absorption into a Living Skin Equivalent Model by MALDI-MSI. Analytical Chemistry 2018, 90, 10031 -10038.
AMA StyleCristina Russo, Neil Brickelbank, Catherine Duckett, Steve Mellor, Stephen Rumbelow, Malcolm R. Clench, Neil Bricklebank. Quantitative Investigation of Terbinafine Hydrochloride Absorption into a Living Skin Equivalent Model by MALDI-MSI. Analytical Chemistry. 2018; 90 (16):10031-10038.
Chicago/Turabian StyleCristina Russo; Neil Brickelbank; Catherine Duckett; Steve Mellor; Stephen Rumbelow; Malcolm R. Clench; Neil Bricklebank. 2018. "Quantitative Investigation of Terbinafine Hydrochloride Absorption into a Living Skin Equivalent Model by MALDI-MSI." Analytical Chemistry 90, no. 16: 10031-10038.
A 3D cell culture is an artificially created environment in which cells are permitted to grow/interact with their surroundings in all three dimensions. Derived from 3D cell culture, organoids are generally small‐scale constructs of cells that are fabricated in the laboratory to serve as 3D representations of in vivo tissues and organs. Due to regulatory, economic and societal issues concerning the use of animals in scientific research it seems clear that the use of 3D cell culture and organoids in for example early stage studies of drug efficacy and toxicity will increase. The combination of such 3D tissue models with mass spectrometry imaging provides a label free methodology for the study of drug absorption/penetration, drug efficacy/toxicity and drug biotransformation. In this article, some of the successes achieved to date and challenges to be overcome before this methodology is more widely adopted are discussed. This article is protected by copyright. All rights reserved
Cristina Russo; Emily E. L. Lewis; Lucy Flint; Malcolm R. Clench. Mass Spectrometry Imaging of 3D Tissue Models. PROTEOMICS 2018, 18, e1700462 .
AMA StyleCristina Russo, Emily E. L. Lewis, Lucy Flint, Malcolm R. Clench. Mass Spectrometry Imaging of 3D Tissue Models. PROTEOMICS. 2018; 18 (14):e1700462.
Chicago/Turabian StyleCristina Russo; Emily E. L. Lewis; Lucy Flint; Malcolm R. Clench. 2018. "Mass Spectrometry Imaging of 3D Tissue Models." PROTEOMICS 18, no. 14: e1700462.
Described is quantitative mass spectrometry imaging (qMSI) methodology for the analysis of lactate and glutamate distributions to delineate heterogeneity between mouse tumor models used to support drug discovery efficacy testing. We evaluate and report on the pre-analysis stabilization methods aimed at improving reproducibility and efficiency of quantitative assessment of endogenous molecules in tissues. Stability experiments demonstrate that optimum stabilization protocols consists of frozen tissue embedding, post tissue-sectioning desiccation and storage of tissue sections at -80°C sealed in vacuum tight containers. Optimized stabilization protocol are used in combination with qMSI methodology for the absolute quantitation of lactate and glutamate in tumors, incorporating the use of two different stable isotope-labeled versions of each analyte and spectral clustering performed on each tissue sections using k-means clustering to allow region-specific pixel by pixel quantitation. Region-specific qMSI was used to screen different tumor models and identify a phenotype that has low lactate heterogeneity to enable accurate measurement of lactate modulation in future drug discovery studies. We conclude that using optimized qMSI protocols it is possible to quantify endogenous metabolites within tumors and that region-specific quantitation can provide valuable insight into tissue heterogeneity and the tumour microenvironment.
John G. Swales; Alex Dexter; Gregory Hamm; Anna Nilsson; Nicole Strittmatter; Filippos Michopoulos; Christopher Hardy; Pablo Morentin-Gutierrez; Martine Mellor; Per E. Andren; Malcolm R. Clench; Josephine Bunch; Susan E. Critchlow; Richard J. A. Goodwin. Quantitation of Endogenous Metabolites in Mouse Tumors Using Mass-Spectrometry Imaging. Analytical Chemistry 2018, 90, 6051 -6058.
AMA StyleJohn G. Swales, Alex Dexter, Gregory Hamm, Anna Nilsson, Nicole Strittmatter, Filippos Michopoulos, Christopher Hardy, Pablo Morentin-Gutierrez, Martine Mellor, Per E. Andren, Malcolm R. Clench, Josephine Bunch, Susan E. Critchlow, Richard J. A. Goodwin. Quantitation of Endogenous Metabolites in Mouse Tumors Using Mass-Spectrometry Imaging. Analytical Chemistry. 2018; 90 (10):6051-6058.
Chicago/Turabian StyleJohn G. Swales; Alex Dexter; Gregory Hamm; Anna Nilsson; Nicole Strittmatter; Filippos Michopoulos; Christopher Hardy; Pablo Morentin-Gutierrez; Martine Mellor; Per E. Andren; Malcolm R. Clench; Josephine Bunch; Susan E. Critchlow; Richard J. A. Goodwin. 2018. "Quantitation of Endogenous Metabolites in Mouse Tumors Using Mass-Spectrometry Imaging." Analytical Chemistry 90, no. 10: 6051-6058.
Examination of the skin barrier repair/wound healing process using a living skin equivalent (LSE) model and matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) to identify lipids directly involved as potential biomarkers. These biomarkers may be used to determine whether an in vivo wound is going to heal for example if infected. An in vitro LSE model was wounded with a scalpel blade and assessed at day 4 post wounding by histology and MALDI-MSI. Samples were sectioned at wound site and were either formalin fixed paraffin embedded (FFPE) for histology or snapped frozen (FF) for MSI analysis. The combination of using an in vitro wounded skin model with MSI allowed the identification of lipids involved in the skin barrier repair/wound healing process. The technique was able to highlight lipids directly in the wound site and distinguish differences in lipid distribution between the epidermis and wound site. This novel method of coupling an in vitro LSE with MSI allowed in depth molecular analysis of the skin barrier repair/wound healing process. The technique allowed the identification of lipids directly involved in the skin barrier repair/wound healing process, indicating these biomarkers may be potentially be used within clinic. These biomarkers will help determine, which stage of the skin barrier repair/wound healing process the wound is in to provide the best treatment. This article is protected by copyright. All rights reserved.
E. E. L. Lewis; M. R. T. Barrett; L. Freeman-Parry; R. A. Bojar; M. R. Clench. Examination of the skin barrier repair/wound healing process using a living skin equivalent model and matrix-assisted laser desorption-ionization-mass spectrometry imaging. International Journal of Cosmetic Science 2018, 40, 148 -156.
AMA StyleE. E. L. Lewis, M. R. T. Barrett, L. Freeman-Parry, R. A. Bojar, M. R. Clench. Examination of the skin barrier repair/wound healing process using a living skin equivalent model and matrix-assisted laser desorption-ionization-mass spectrometry imaging. International Journal of Cosmetic Science. 2018; 40 (2):148-156.
Chicago/Turabian StyleE. E. L. Lewis; M. R. T. Barrett; L. Freeman-Parry; R. A. Bojar; M. R. Clench. 2018. "Examination of the skin barrier repair/wound healing process using a living skin equivalent model and matrix-assisted laser desorption-ionization-mass spectrometry imaging." International Journal of Cosmetic Science 40, no. 2: 148-156.
Introduction: The distinction of papillary thyroid carcinomas from benign thyroid lesions has important implication for clinical management. Classification based on histopathological features can be supported by molecular biomarkers, including lipidomic signatures, identified with the use of high-throughput mass spectrometry techniques. Formalin fixation is a standard procedure for stabilization and preservation of tissue samples, therefore this type of samples constitute highly valuable source of clinical material for retrospective molecular studies. In this study we used mass spectrometry imaging to detect lipids discriminating papillary cancer from not cancerous thyroid directly in formalin-fixed tissue sections. Material and methods: For this purpose imaging and profiling of lipids present in non-malignant and cancerous thyroid tissue specimens were conducted. High resolution MALDI-Q-Ion Mobility-TOF-MS technique was used for lipidomic analysis of formalin fixed thyroid tissue samples. Lipids were identified by the comparison of the exact molecular masses and fragmentation pathways of the protonated molecule ions, recorded during the MS/MS experiments, with LIPID MAPS database. Results: Several phosphatidylcholines (32:0, 32:1, 34:1 and 36:3), sphingomyelins (34:1 and 36:1) and phosphatidic acids (36:2 and 36:3) were detected and their abundances were significantly higher in cancerous tissue compared to non-cancerous tissue. The same lipid species were detected in formalin-fixed as in fresh-frozen tissue, but [M + Na]+ ions were the most abundant in formalin fixed whereas [M + K]+ ions were predominant in fresh tissue. Conclusions: Our results prove the viability of MALDI-MSI for analysis of lipid distribution directly in formalin-fixed tissue, and the potential for their use in the classification of thyroid diseases. Wstęp: Rozróżnienie brodawkowatych raków tarczycy od łagodnych zmian tarczycy ma istotny wpływ na postepowanie kliniczne. Klasyfikacja w oparciu o cechy histopatologiczne może być wspomagana biomarkerami molekularnymi, w tym składnikami lipidowymi, zidentyfikowanymi przy użyciu wysokorozdzielczych technik spektrometrii masowej. Utrwalanie tkanki w formalinie jest standardową procedurą stabilizacji i konserwacji tkanek, które stanowią cenne źródło materiału klinicznego w retrospektywnych badaniach molekularnych. W prezentowanych badaniach wykorzystano obrazowanie molekularne metodą MALDI w celu wykrycia składników lipidomu, obecnych w tkankach utrwalonych w formalinie, różnicujących raka brodawkowatego od tarczycy nienowotworowej. Materiał i metody: W tym celu przeprowadzono obrazowanie i profilowanie lipidów obecnych w próbkach tkanek tarczycy zmienionej nowotworowo i nienowotworowej. Do analizy lipidomicznej tkanek tarczycy utrwalonych w formalinie zastosowano wysokorozdzielczą technikę MALDI-Q-Ion Mobility-TOF-MS. Składniki lipidowe zostały zidentyfikowane przez porównanie uzyskanych dokładnych mas cząsteczkowych i szlaków fragmentacji jonów protonowanych, zarejestrowanych podczas eksperymentów MS/MS, z bazą danych LIPID MAPS. Wyniki: W oparciu o przeprowadzone analizy wykryto lipidy należące do klas fosfatydylocholin (32:0, 32:1, 34:1 i 36:3), sfingomielin (34:1 i 36:1) i kwasów fosfatydowych (36:2 i 36:3), a ich ilość była znacząco wyższa w tkance nowotworowej w porównaniu z tarczycą nienowotworową. Te same składniki lipidowe wykryto w preparatach utrwalonych w formalinie, jak i w tkance świeżo mrożonej, przy czym jony [M + Na]+ obecne były w przewadze w tkance utrwalonej w formalinie, podczas gdy w świeżej tkance przeważały jony [M + K]+. Wnioski: Prezentowane wyniki potwierdzają możliwości wykorzystania techniki MALDI-MSI w analizie dystrybucji lipidów bezpośrednio w tkankach stabilizowanych formaliną oraz dają nadzieję na potencjalne wykorzystanie ich w klasyfikacji chorób tarczycy.
Anna Wojakowska; Laura M. Cole; Mykola Chekan; Katarzyna Bednarczyk; Magdalena Maksymiak; Malgorzata Oczko-Wojciechowska; Barbara Jarząb; Malcolm R. Clench; Joanna Polanska; Monika Pietrowska; Piotr Widlak. Odróżnienie brodawkowatego raka tarczycy od tkanki nienowotworowej w oparciu o profilowanie lipidów metodą MALDI-MSI. Endokrynologia Polska 2018, 69, 2 -8.
AMA StyleAnna Wojakowska, Laura M. Cole, Mykola Chekan, Katarzyna Bednarczyk, Magdalena Maksymiak, Malgorzata Oczko-Wojciechowska, Barbara Jarząb, Malcolm R. Clench, Joanna Polanska, Monika Pietrowska, Piotr Widlak. Odróżnienie brodawkowatego raka tarczycy od tkanki nienowotworowej w oparciu o profilowanie lipidów metodą MALDI-MSI. Endokrynologia Polska. 2018; 69 (1):2-8.
Chicago/Turabian StyleAnna Wojakowska; Laura M. Cole; Mykola Chekan; Katarzyna Bednarczyk; Magdalena Maksymiak; Malgorzata Oczko-Wojciechowska; Barbara Jarząb; Malcolm R. Clench; Joanna Polanska; Monika Pietrowska; Piotr Widlak. 2018. "Odróżnienie brodawkowatego raka tarczycy od tkanki nienowotworowej w oparciu o profilowanie lipidów metodą MALDI-MSI." Endokrynologia Polska 69, no. 1: 2-8.
MALDI-mass spectrometry imaging (MALDI-MSI) has been shown to allow the study of protein distribution and identification directly within formalin-fixed paraffin-embedded (FFPE) tissue sections. However, direct protein identification from tissue sections remains challenging due to signal interferences and/or existing post-translational or other chemical modifications. The use of antigen retrieval (AR) has been demonstrated for unlocking proteins prior to in situ enzymatic digestion and MALDI-MSI analysis of FFPE tissue sections. In the work reported here, the identification of proline oxidation, which may occur when performing the AR protocol, is described. This facilitated and considerably increased the number of identified peptides when adding proline oxidation as a variable modification to the MASCOT search criteria. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann.
Marie-Claude Djidja; Emmanuelle Claude; Peter Scriven; David W. Allen; Vikki A. Carolan; Malcolm R. Clench. Antigen retrieval prior to on-tissue digestion of formalin-fixed paraffin-embedded tumour tissue sections yields oxidation of proline residues. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 2017, 1865, 901 -906.
AMA StyleMarie-Claude Djidja, Emmanuelle Claude, Peter Scriven, David W. Allen, Vikki A. Carolan, Malcolm R. Clench. Antigen retrieval prior to on-tissue digestion of formalin-fixed paraffin-embedded tumour tissue sections yields oxidation of proline residues. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 2017; 1865 (7):901-906.
Chicago/Turabian StyleMarie-Claude Djidja; Emmanuelle Claude; Peter Scriven; David W. Allen; Vikki A. Carolan; Malcolm R. Clench. 2017. "Antigen retrieval prior to on-tissue digestion of formalin-fixed paraffin-embedded tumour tissue sections yields oxidation of proline residues." Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 1865, no. 7: 901-906.
This study investigates the identity of two unexpected arsenic species found separately in a number of urine samples sent to the Health and Safety Executive’s Health and Safety Laboratory for arsenic speciation (arsenobetaine, AB; arsenite, As3+; arsenate, As5+; monomethylarsonic acid, MMA5+; and dimethylarsinic acid, DMA5+). Micro liquid chromatography coupled to inductively coupled plasma mass spectrometry (µLC-ICP-MS) and electrospray time of flight tandem mass spectrometry (ESI-QqTOF-MS/MS) were used to identify the two arsenic peaks by comparison to several characterized arsenicals: arsenocholine, AC; trimethyl arsine oxide, TMAO; dimethylarsenoacetate, DMAA; dimethylarsenoethanol, DMAE; thio-dimethylarsinate, thio-DMA; thio-dimethylarsenoacetate, thio-DMAA and thio-dimethylarsenoethanol, thio-DMAE. The results from both the ICP-MS and ESI-QqTOF-MS/MS investigations indicate that the unexpected arsenic species termed peak 1 was thio-DMA. While the unexpected arsenic species termed peak 2 has yet to be identified, this investigation shows that it was not AC, TMAO, DMAA, DMAE, thio-DMA, thio-DMAA or thio-DMAE. This study demonstrates the incidence of unexpected arsenic species in both routine and non-routine urine samples from both workers and hospital patients.
Elizabeth Leese; Malcolm Clench; Jackie Morton; Philip H.E. Gardiner; Vikki A. Carolan. The Investigation of Unexpected Arsenic Compounds Observed in Routine Biological Monitoring Urinary Speciation Analysis. Toxics 2017, 5, 12 .
AMA StyleElizabeth Leese, Malcolm Clench, Jackie Morton, Philip H.E. Gardiner, Vikki A. Carolan. The Investigation of Unexpected Arsenic Compounds Observed in Routine Biological Monitoring Urinary Speciation Analysis. Toxics. 2017; 5 (2):12.
Chicago/Turabian StyleElizabeth Leese; Malcolm Clench; Jackie Morton; Philip H.E. Gardiner; Vikki A. Carolan. 2017. "The Investigation of Unexpected Arsenic Compounds Observed in Routine Biological Monitoring Urinary Speciation Analysis." Toxics 5, no. 2: 12.