Towards monitoring real-time cellular response using an integrated microfluidics-matrix assisted laser desorption ionisation/nanoelectrospray ionisation-ion mobility-mass spectrometry platform.

The combination of microfluidic cell trapping devices with ion mobility-mass spectrometry offers the potential for elucidating in real time the dynamic responses of small populations of cells to paracrine signals, changes in metabolite levels and delivery of drugs and toxins. Preliminary experiments examining peptides in methanol and recording the interactions of yeast and Jurkat cells with their superfusate have identified instrumental set-up and control parameters and online desalting procedures. Numerous initial experiments demonstrate and validate this new instrumental platform. Future outlooks and potential applications are addressed, specifically how this instrumentation may be used for fully automated systems biology studies of the significantly interdependent, dynamic internal workings of cellular metabolic and signalling pathways.

[1]  S. Valentine,et al.  High-order structure and dissociation of gaseous peptide aggregates that are hidden in mass spectra , 1998, Journal of the American Society for Mass Spectrometry.

[2]  Kermit K. Murray,et al.  Development of an efficient on-chip digestion system for protein analysis using MALDI-TOF MS. , 2009, The Analyst.

[3]  T. Rejtar,et al.  High-speed, high-resolution monolithic capillary LC-MALDI MS using an off-line continuous deposition interface for proteomic analysis. , 2005, Analytical chemistry.

[4]  S. E. Eklund,et al.  Metabolic Discrimination of Select List Agents by Monitoring Cellular Responses in a Multianalyte Microphysiometer , 2009, Sensors.

[5]  N. Anderson,et al.  The Human Plasma Proteome , 2002, Molecular & Cellular Proteomics.

[6]  Nicholas A. Cellar,et al.  A capillary-PDMS hybrid chip for separations-based sensing of neurotransmitters in vivo. , 2006, Lab on a chip.

[7]  E. W. McDaniel,et al.  Collision phenomena in ionized gases , 1964 .

[8]  J. McLean.  The mass-mobility correlation redux: The conformational landscape of anhydrous biomolecules , 2009, Journal of the American Society for Mass Spectrometry.

[9]  Nicholas A. Cellar,et al.  Microfluidic chip for low-flow push-pull perfusion sampling in vivo with on-line analysis of amino acids. , 2005, Analytical chemistry.

[10]  Gwo-Bin Lee,et al.  Micro devices integrated with microchannels and electrospray nozzles using PDMS casting techniques , 2002 .

[11]  Ales Prokop,et al.  A microphysiometer for simultaneous measurement of changes in extracellular glucose, lactate, oxygen, and acidification rate. , 2004, Analytical chemistry.

[12]  E. A. Lima,et al.  A microfluidic device to confine a single cardiac myocyte in a sub-nanoliter volume on planar microelectrodes for extracellular potential recordings. , 2004, Lab on a chip.

[13]  Jody C. May,et al.  Dual source ion mobility-mass spectrometer for direct comparison of electrospray ionization and MALDI collision cross section measurements. , 2010, Analytical chemistry.

[14]  M. Moon,et al.  Nanoflow LC/ion mobility/CID/TOF for proteomics: analysis of a human urinary proteome. , 2003, Journal of proteome research.

[15]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[16]  F. McLafferty,et al.  Attomole Protein Characterization by Capillary Electrophoresis-Mass Spectrometry , 1996, Science.

[17]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[18]  Richard D. Smith,et al.  ON-LINE MICRODIALYSIS DESALTING FOR ELECTROSPRAY IONIZATION-MASS SPECTROMETRY OF PROTEINS AND PEPTIDES , 1996 .

[19]  A. Edefonti,et al.  Shiga Toxins Present in the Gut and in the Polymorphonuclear Leukocytes Circulating in the Blood of Children with Hemolytic-Uremic Syndrome , 2006, Journal of Clinical Microbiology.

[20]  S. E. Eklund,et al.  Multianalyte microphysiometry as a tool in metabolomics and systems biology , 2006 .

[21]  E. A. Lima,et al.  Thin-film IrOx pH microelectrode for microfluidic-based microsystems. , 2005, Biosensors & bioelectronics.

[22]  D. Clemmer,et al.  Gas-phase separations of electrosprayed peptide libraries. , 1999, Analytical chemistry.

[23]  John P Wikswo,et al.  Microfluidic platform for real-time signaling analysis of multiple single T cells in parallel. , 2008, Lab on a chip.

[24]  V. Longo,et al.  The chronological life span of Saccharomyces cerevisiae , 2003, Methods in molecular biology.

[25]  Microfluidic device to confine single cardiac myocytes in sub-nanoliter volumes for extracellular pH measurements , 2008 .

[26]  S. Huang,et al.  Dimensions of systems biology. , 2006, Reviews of physiology, biochemistry and pharmacology.

[27]  Hod Lipson,et al.  Automated reverse engineering of nonlinear dynamical systems , 2007, Proceedings of the National Academy of Sciences.

[28]  R. Aebersold,et al.  Mass spectrometry-based proteomics , 2003, Nature.

[29]  Larissa S Fenn,et al.  Biomolecular structural separations by ion mobility–mass spectrometry , 2008, Analytical and bioanalytical chemistry.

[30]  Amanda Clare,et al.  An ontology for a Robot Scientist , 2006, ISMB.

[31]  L. Locascio,et al.  Integrated plastic microfluidic devices with ESI-MS for drug screening and residue analysis. , 2001, Analytical chemistry.

[32]  J P Wikswo,et al.  Engineering challenges of BioNEMS: the integration of microfluidics, micro- and nanodevices, models and external control for systems biology. , 2006, IEE proceedings. Nanobiotechnology.

[33]  Markus Stoeckli,et al.  MALDI mass spectrometric imaging of biological tissue sections , 2005, Mechanisms of Ageing and Development.

[34]  Larissa S Fenn,et al.  Simultaneous glycoproteomics on the basis of structure using ion mobility-mass spectrometry. , 2009, Molecular bioSystems.

[35]  G. Whitesides,et al.  Microfluidic devices fabricated in Poly(dimethylsiloxane) for biological studies , 2003, Electrophoresis.

[36]  Frantisek Foret,et al.  Immobilized microfluidic enzymatic reactors , 2004, Electrophoresis.

[37]  D. Jed Harrison,et al.  Analytical microdevices for mass spectrometry , 2000 .

[38]  M. Hennion.  Solid-phase extraction: method development, sorbents, and coupling with liquid chromatography. , 1999, Journal of chromatography. A.

[39]  Stephen Naylor,et al.  Mapping the human plasma proteome by SCX-LC-IMS-MS , 2007, Journal of the American Society for Mass Spectrometry.

[40]  J. Sweedler,et al.  Online microdialysis-dynamic nanoelectrospray ionization-mass spectrometry for monitoring neuropeptide secretion. , 2005, Journal of mass spectrometry : JMS.

[41]  E. W. McDaniel,et al.  The transport of slow ions in gases: Experiment, theory, and applications , 1984 .

[42]  Michael G. Roper,et al.  Microfluidic chip for continuous monitoring of hormone secretion from live cells using an electrophoresis-based immunoassay. , 2003, Analytical chemistry.

[43]  R. Caprioli,et al.  Micro-electrospray mass spectrometry: Ultra-high-sensitivity analysis of peptides and proteins , 1994, Journal of the American Society for Mass Spectrometry.

[44]  I. Ges,et al.  On-chip acidification rate measurements from single cardiac cells confined in sub-nanoliter volumes , 2008, Biomedical microdevices.

[45]  J. Wikswo,et al.  Quantum Dot Probes for Monitoring Dynamic Cellular Response: Reporters of T Cell Activation , 2006, IEEE Transactions on NanoBioscience.

[46]  Hod Lipson,et al.  Resilient Machines Through Continuous Self-Modeling , 2006, Science.

[47]  R. Aebersold,et al.  Microfabricated polymer devices for automated sample delivery of peptides for analysis by electrospray ionization tandem mass spectrometry. , 1999, Analytical chemistry.

[48]  Robert T Kennedy,et al.  Perfusion and chemical monitoring of living cells on a microfluidic chip. , 2005, Lab on a chip.

[49]  J. Ramsey,et al.  Fully integrated glass microfluidic device for performing high-efficiency capillary electrophoresis and electrospray ionization mass spectrometry. , 2008, Analytical chemistry.

[50]  I. Ges,et al.  Differential pH measurements of metabolic cellular activity in nl culture volumes using microfabricated iridium oxide electrodes. , 2007, Biosensors & bioelectronics.

[51]  Ken E. Whelan,et al.  The Automation of Science , 2009, Science.

[52]  Edward A. Mason,et al.  Transport Properties of Ions in Gases: MASON:TRANSPORT PROPERTIES O-BK , 2005 .

[53]  S. E. Eklund,et al.  Modification of the Cytosensor™ microphysiometer to simultaneously measure extracellular acidification and oxygen consumption rates , 2003 .

[54]  Donald Wlodkowic,et al.  Microfluidic single cell arrays to interrogate signalling dynamics of individual, patient-derived hematopoietic stem cells. , 2009, Lab on a chip.

[55]  Larissa S Fenn,et al.  Characterizing ion mobility-mass spectrometry conformation space for the analysis of complex biological samples , 2009, Analytical and bioanalytical chemistry.

[56]  E. W. McDaniel,et al.  Transport Properties of Ions in Gases , 1988 .

[57]  Hod Lipson,et al.  Distilling Free-Form Natural Laws from Experimental Data , 2009, Science.