LESA MS Imaging of Heat-Preserved and Frozen Tissue: Benefits of Multistep Static FAIMS.

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.

[1]  H. Cooper,et al.  Native mass spectrometry imaging of intact proteins and protein complexes in thin tissue sections , 2017, International Journal of Mass Spectrometry.

[2]  L. Eberlin,et al.  Desorption Electrospray Ionization Mass Spectrometry Imaging of Proteins Directly from Biological Tissue Sections. , 2018, Analytical chemistry.

[3]  H. Cooper,et al.  Ambient surface mass spectrometry-ion mobility spectrometry of intact proteins. , 2018, Current opinion in chemical biology.

[4]  J. Griffin,et al.  Liquid Extraction Surface Analysis Mass Spectrometry Method for Identifying the Presence and Severity of Nonalcoholic Fatty Liver Disease. , 2017, Analytical chemistry.

[5]  L. Eberlin,et al.  Ambient Ionization and FAIMS Mass Spectrometry for Enhanced Imaging of Multiply Charged Molecular Ions in Biological Tissues. , 2016, Analytical chemistry.

[6]  Josephine Bunch,et al.  SpectralAnalysis: Software for the Masses. , 2016, Analytical chemistry.

[7]  H. Cooper,et al.  LESA FAIMS Mass Spectrometry for the Spatial Profiling of Proteins from Tissue. , 2016, Analytical chemistry.

[8]  H. Cooper.  To What Extent is FAIMS Beneficial in the Analysis of Proteins? , 2016, Journal of The American Society for Mass Spectrometry.

[9]  H. Cooper,et al.  Direct Tissue Profiling of Protein Complexes: Toward Native Mass Spectrometry Imaging. , 2016, Analytical chemistry.

[10]  Peter J H Webborn,et al.  Mapping drug distribution in brain tissue using liquid extraction surface analysis mass spectrometry imaging. , 2015, Analytical chemistry.

[11]  Harish Shankaran,et al.  Investigating nephrotoxicity of polymyxin derivatives by mapping renal distribution using mass spectrometry imaging. , 2015, Chemical research in toxicology.

[12]  Elizabeth C. Randall,et al.  Liquid Extraction Surface Analysis Mass Spectrometry Coupled with Field Asymmetric Waveform Ion Mobility Spectrometry for Analysis of Intact Proteins from Biological Substrates. , 2015, Analytical chemistry.

[13]  H. Cooper,et al.  Native Liquid Extraction Surface Analysis Mass Spectrometry: Analysis of Noncovalent Protein Complexes Directly from Dried Substrates , 2015, Journal of The American Society for Mass Spectrometry.

[14]  J. Vance.  Phospholipid Synthesis and Transport in Mammalian Cells , 2015, Traffic.

[15]  Elizabeth C. Randall,et al.  Direct analysis of intact proteins from Escherichia coli colonies by liquid extraction surface analysis mass spectrometry. , 2014, Analytical chemistry.

[16]  A. Shvartsburg.  Ultrahigh-resolution differential ion mobility separations of conformers for proteins above 10 kDa: onset of dipole alignment? , 2014, Analytical chemistry.

[17]  H. Cooper,et al.  Top-Down and Bottom-Up Identification of Proteins by Liquid Extraction Surface Analysis Mass Spectrometry of Healthy and Diseased Human Liver Tissue , 2014, Journal of The American Society for Mass Spectrometry.

[18]  Peter J H Webborn,et al.  Mass spectrometry imaging of cassette-dosed drugs for higher throughput pharmacokinetic and biodistribution analysis. , 2014, Analytical chemistry.

[19]  H. Cooper,et al.  Compound heterozygotes and beta‐thalassemia: Top‐down mass spectrometry for detection of hemoglobinopathies , 2014, Proteomics.

[20]  T. Porta,et al.  Gas-phase separation of drugs and metabolites using modifier-assisted differential ion mobility spectrometry hyphenated to liquid extraction surface analysis and mass spectrometry. , 2013, Analytical chemistry.

[21]  F. Betsou,et al.  Lithostatine concentration is not useful for assessing the preanalytical variations in biobanked urine samples. , 2013, Biopreservation and biobanking.

[22]  Richard D. Smith,et al.  Separation of protein conformers by differential ion mobility in hydrogen-rich gases. , 2013, Analytical chemistry.

[23]  H. Cooper,et al.  Probing the Complementarity of FAIMS and Strong Cation Exchange Chromatography in Shotgun Proteomics , 2013, Journal of the American Society for Mass Spectrometry.

[24]  Richard D. Smith,et al.  High-resolution differential ion mobility spectrometry of a protein. , 2013, Analytical chemistry.

[25]  H. Cooper,et al.  Top-Down Proteomics and Direct Surface Sampling of Neonatal Dried Blood Spots: Diagnosis of Unknown Hemoglobin Variants , 2012, Journal of The American Society for Mass Spectrometry.

[26]  Richard D. Smith,et al.  Protein analyses using differential ion mobility microchips with mass spectrometry. , 2012, Analytical chemistry.

[27]  C. Mackay,et al.  Conductive carbon tape used for support and mounting of both whole animal and fragile heat-treated tissue sections for MALDI MS imaging and quantitation. , 2012, Journal of proteomics.

[28]  K. Kultima,et al.  Biomarkers of disease and post-mortem changes - Heat stabilization, a necessary tool for measurement of protein regulation. , 2011, Journal of proteomics.

[29]  K. Gardiner,et al.  Preserving protein profiles in tissue samples: Differing outcomes with and without heat stabilization , 2011, Journal of Neuroscience Methods.

[30]  H. Cooper,et al.  Hemoglobin Variant Analysis via Direct Surface Sampling of Dried Blood Spots Coupled with High-Resolution Mass Spectrometry , 2011, Analytical chemistry.

[31]  F. Fernández,et al.  Enhanced direct ambient analysis by differential mobility-filtered desorption electrospray ionization-mass spectrometry. , 2010, Analytical chemistry.

[32]  Mats Borén,et al.  Stopping the clock on proteomic degradation by heat treatment at the point of tissue excision , 2010, Proteomics.

[33]  Marcus Svensson,et al.  Heat stabilization of the tissue proteome: a new technology for improved proteomics. , 2009, Journal of proteome research.

[34]  R. Caprioli,et al.  Direct tissue analysis by matrix-assisted laser desorption ionization mass spectrometry: application to kidney biology. , 2007, Seminars in nephrology.

[35]  Richard M Caprioli,et al.  Molecular imaging of thin mammalian tissue sections by mass spectrometry. , 2006, Current opinion in biotechnology.

[36]  Richard D. Smith,et al.  Characterizing the structures and folding of free proteins using 2-D gas-phase separations: observation of multiple unfolded conformers. , 2006, Analytical chemistry.

[37]  D. Barnett,et al.  Combining HD exchange and ESI-FAIMS-MS for detecting gas-phase conformers of equine cytochrome c , 2005 .

[38]  Pierre Thibault,et al.  Improvement in peptide detection for proteomics analyses using NanoLC-MS and high-field asymmetry waveform ion mobility mass spectrometry. , 2005, Analytical chemistry.

[39]  S. Radford,et al.  Separation of β2-microglobulin conformers by high-field asymmetric waveform ion mobility spectrometry (FAIMS) coupled to electrospray ionisation mass spectrometry , 2004 .

[40]  G. V. Van Berkel,et al.  Thin-layer chromatography and electrospray mass spectrometry coupled using a surface sampling probe. , 2002, Analytical chemistry.

[41]  R. Purves,et al.  Electrospray ionization high-field asymmetric waveform ion mobility spectrometry-mass spectrometry. , 1999, Analytical chemistry.

[42]  R. Purves,et al.  MASS SPECTROMETRIC CHARACTERIZATION OF A HIGH-FIELD ASYMMETRIC WAVEFORM ION MOBILITY SPECTROMETER , 1998 .