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    Flow Cytometry

    Flow Cytometry: a laser- or impedance-based, biophysical technology employed in cell counting, cell sorting, biomarker detection and protein engineering, by suspending cells in a stream of fluid and passing them through an electronic detection apparatus. A flow cytometer allows simultaneous multiparametric analysis of the physical and chemical characteristics of up to thousands of particles per second. Flow cytometry is routinely used in the diagnosis of health disorders, especially blood cancers, but has many other applications in basic research, clinical practice and clinical trials.

    Webinars

    • NOV 21, 2019 | 8:00 AM
      How to maximize tumor microenvironment analysis from limited patient samples
      How to maximize tumor microenvironment analysis from limited patient...
      DATE: November 21, 2019TIME: 8:00am PSTA complex interaction among heterogenous immune and tumor cells within the tumorous tissue not only influences tumor progression, but can also sha...
      Speaker: César Evaristo, PhD
      Sponsored By: Miltenyi Biotec
      5 2
      NOV 18, 2019 | 7:00 AM
      Protect your cells with proper pipetting - How liquid handling influences your cell culture work
      Protect your cells with proper pipetting - How liquid handling...
      DATE: November 18, 2019TIME: 7:00am PST, 11:00am EST, 4:00pm CEWT How often do you pipette in your cell culture lab every day? Usually, we do it so often that we tend stop th...
      Speaker: Nadine Mellies, PhD , Katja Baesler, PhD
      Sponsored By: Eppendorf
      22 2
      NOV 07, 2019 | 10:00 AM
      Akoya Biosciences Webinar Series: Validating Multiplexed Antibody Panels for Pancreatic Research Applications
      Akoya Biosciences Webinar Series: Validating Multiplexed Antibody...
      DATE: November 7, 2019TIME: 10:00am PST, 1:00pm EST Studying the pathogenesis of diabetes requires detailed analysis of the pancreatic islet microenvironment and its numerous c...
      Speaker: Diane Saunders, PhD , Grady Carlson, PhD
      Sponsored By: Akoya Biosciences
      5
      OCT 31, 2019 | 9:00 AM
      Investigation of Endothelial Cell Viability and Growth on 3D printed GelMa Vascular Networks
      Investigation of Endothelial Cell Viability and Growth on 3D printed...
      DATE: October 31, 2019TIME: 9:00am PDT, 12:00pm EDT INTRODUCTION: A major limitation for the development of 3D engineered tissues is the absence of viable and perfusable...
      Speaker: Chrisoula Chappell
      Sponsored By: Thermo Fisher Scientific/Gibco
      8
      OCT 24, 2019 | 10:30 AM
      Supercritical fluid chromatography of polar biomolecules at the analytical and preparative scale: evaluation of global conformational changes in peptides and proteins
      Supercritical fluid chromatography of polar biomolecules at the...
      Supercritical fluid chromatography (SFC) has historically been very effective for exacting difficult chiral separations, with an unmatched power in terms of enabling highly efficient enantios...
      Speaker: Raffeal Bennett, PhD
      Presented at: Analytical Chemistry Virtual Event Series 2019
      2
      OCT 24, 2019 | 6:00 AM
      Multidimension Chromatography; a Disruptive Technology - A Netflix Effect in the World of Chromatography!
      Multidimension Chromatography; a Disruptive Technology - A Netflix...
      Chromatography is the first choice in separation technique for a wide selection of applications, across many fields, ranging from simple to highly complex extracts. The main rationale for it...
      Speaker: Claude Mallet, PhD
      Presented at: Analytical Chemistry Virtual Event Series 2019
      2
      OCT 22, 2019 | 10:00 AM
      Demystifying Analytical Validation While Onboarding NGS Tests
      Demystifying Analytical Validation While Onboarding NGS Tests
      DATE: October 22, 2019TIME: 10:00am PDTNext-generation sequencing (NGS) is quickly becoming the go-to tool for testing labs looking to bring on additional clinically relevant tests requ...
      Speaker: Geoffrey Bien , Leah Ames, MS, MLS (ASCP) SCYM
      Sponsored By: Thermo Fisher Scientific
      5 3
      SEP 24, 2019 | 8:00 AM
      Interrogating 3D spheroid versus 2D monolayer cell models in drug discovery using cell health microplate assays
      Interrogating 3D spheroid versus 2D monolayer cell models in drug...
      DATE: September 24, 2019TIME: 7:00am PDTHigh throughput screening (HTS) is an effective method for identifying putative active compounds for therapeutics. Assays that evaluate changes i...
      Speaker: Dr. Adyary Fallarero, Ph.D. Adjunct Professor/Docent in Pharmaceutical Biology , Dr. Leticia Montoya, Ph.D., Chemistry and Biochemistry
      Sponsored By: Thermo Fisher Scientific
      9
      AUG 27, 2019 | 9:00 AM
      Akoya Biosciences Webinar Series:  Comprehensive Meta-analysis of Biomarker Technologies for Predictive Response of PD-1/PD-L1 Checkpoint Therapies
      Akoya Biosciences Webinar Series: Comprehensive Meta-analysis...
      DATE: August 27, 2019 TIME: 9:00am PDT, 12:00pm EDT Immunotherapies targeting PD-1 or PD-L1 have proven remarkably effective for treating cancer in some patients, with considerabl...
      Speaker: Cliff Hoyt, PhD , Steve Lu
      Sponsored By: Akoya Biosciences
      5
      JUL 24, 2019 | 8:00 AM
      Establishing a robust 37-marker mass cytometry assay for deep single-cell immune profiling of whole blood
      Establishing a robust 37-marker mass cytometry assay for deep...
      DATE: July 24, 2019TIMEL 8:00am PT Mass cytometry (CyTOF®) is a powerful approach to characterize the immune composition of complex biological specimens and gain insights in...
      Speaker: Adeeb Rahman, PhD , Mona Al-Maarri, PhD
      Sponsored By: Miltenyi Biotec
      8
      JUN 05, 2019 | 5:00 PM
      Everyday culture practice - Improving reproducibility in cell culture
      Everyday culture practice - Improving reproducibility in cell...
      DATE: June 5, 2019TIME: 8:00am PDT, 11:00am EDT, 5:00pm CEST Eukaryotic cell cultures respond to the most subtle influence. Apart from the risk of contamination, minimal chan...
      Speaker: Jessica Wagener, PhD
      Sponsored By: Eppendorf
      23 2
      MAY 16, 2019 | 12:00 PM
      Isolation and expansion of mesenchymal stromal or stem cells (MSCs) for translational and clinical research
      Isolation and expansion of mesenchymal stromal or stem cells (MSCs)...
      DATE: May 16, 2019TIME: 12:00 PM PDT Mesenchymal stem or stromal cells (MSCs) are a promising resource for cell-based therapies and are being tested in already registered phase...
      Speaker: Kathrin Godthardt, B.Sc.
      Sponsored By: Miltenyi Biotec
      11
      MAY 16, 2019 | 4:00 PM
      Towards Standardized Workflows - A focus on microbiome insights
      Towards Standardized Workflows - A focus on microbiome insights
      DATE: May 16, 2019TIME: 7:00am PDT, 10:00am EDT, 4:00pm CEST The emergence of NGS is revolutionizing the microbiological sciences and transforming medicine. Deep sequencing has...
      Speaker: Rita Colwell, PhD , Christiane Honisch, PhD
      Sponsored By: Eppendorf
      10 2
      MAR 05, 2019 | 10:00 AM
      Comprehensive Capture of Human Neuropathology by Multiplexed Ion Beam Imaging (MIBI)
      Comprehensive Capture of Human Neuropathology by Multiplexed Ion...
      DATE: March 5, 2019TIME:  10:00am PSTSingle cell analysis, starting with the earliest low parameter fluorescent experiments, helped define the major cell subsets of human ce...
      Speaker: Sean Bendall, PhD
      Sponsored By: IONpath
      2 1
      DEC 05, 2018 | 8:00 PM
      The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary Tumors and in Circulation
      The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary...
      DATE: December 05, 2018TIME:  08:00 PSTWhat is desperately needed are methods to detect cells that cause metastasis–known as cancer stem cells (CSC), a subpopulation of...
      Speaker: Bruno Sainz, PhD
      Sponsored By: Thermo Fisher Scientific
      5 3
      DEC 05, 2018 | 7:00 AM
      The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary Tumors and in Circulation
      The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary...
      DATE: December 05, 2018TIME:  07:00 PSTWhat is desperately needed are methods to detect cells that cause metastasis–known as cancer stem cells (CSC), a subpopulation of...
      Speaker: Bruno Sainz, PhD
      Sponsored By: Thermo Fisher Scientific
      7
      NOV 29, 2018 | 9:00 AM
      An overview of primary immune deficiency disorders (PID) and laboratory testing for PID
      An overview of primary immune deficiency disorders (PID) and laboratory...
      DATE: November 29, 2018TIME:  9:00AM PDTAn overview of primary immune deficiency disorders (PID) and laboratory testing for PIDPrimary immune deficiencies (PIDs), oth...
      Speaker: Vijaya Knight, MD, PhD, (D)ABMLI
      Sponsored By: Binding Site
      10
      NOV 15, 2018 | 10:00 AM
      Multiplex fluorescence immunohistochemistry using the Ultivue InSituPlex platform on the Leica Biosystems BOND RX
      Multiplex fluorescence immunohistochemistry using the Ultivue...
      DATE:  November 15, 2018TIME:  10:00am PT, 1:00pm ET Multiplex fluorescence immunohistochemistry offers a window into the biology of human disease, enabling the ana...
      Speaker: Traci DeGeer, BS, HT (ASCP) HTL, QIHC , Alexander Klimowicz
      Sponsored By: Leica Biosystems
      6 1
      NOV 07, 2018 | 8:00 PM
      Analysis of Surface Antigens on Exosomes
      Analysis of Surface Antigens on Exosomes
      DATE: November 7, 2018TIME: 8:00PM PSTExosomes have been shown to have significant roles in cancer including disease progression acting in the tumor micro-environment, metasta...
      Speaker: Steve McClellan BS, MT, SCYM (ASCP)CM
      Sponsored By: Thermo Fisher Scientific
      3 1
      NOV 07, 2018 | 7:00 AM
      Analysis of Surface Antigens on Exosomes
      Analysis of Surface Antigens on Exosomes
      DATE: November 7, 2018TIME: 7:00AM PDTExosomes have been shown to have significant roles in cancer including disease progression acting in the tumor micro-environment, metastasis a...
      Speaker: Steve McClellan BS, MT, SCYM (ASCP)CM
      Sponsored By: Thermo Fisher Scientific
      11 1
      NOV 01, 2018 | 12:00 AM
      Real-time Live-cell Analysis for Immunologists
      Real-time Live-cell Analysis for Immunologists
      Deriving deeper biological insight & improving productivity in immune-cell biologyThe most common in vitro methods for immunologists to analyze cells of the immune system are flow cyt...
      Speaker: Del Trezise, PhD
      Presented at: Dimension of Time: From Culture to Assay
      2
      OCT 25, 2018 | 12:00 PM
      Through the Looking Glass, and What Amino Acids Found There
      Through the Looking Glass, and What Amino Acids Found There
      Analytical chemistry can take you to strange places - for me it has been to the bottom of a quarry, being covered in dirt, excavating sesame-seed sized fossils that allow us to work out when...
      Speaker: Kirsty Penkman, PhD
      Presented at: Analytical Chemistry Virtual Event Series 2018
      1
      OCT 25, 2018 | 10:30 AM
      Trash or Treasure: Optimizing Titration Electrodes and Consumables
      Trash or Treasure: Optimizing Titration Electrodes and Consumables
      Everyone that uses titration in their lab knows how simple and fast the technique can be.  However, uncertainty around when to replace electrodes creates confusion. Reagent quality, tubi...
      Speaker: Jessica McVay , Lori Spafford
      Presented at: Analytical Chemistry Virtual Event Series 2018
      Sponsored By: Metrohm USA
      1
      OCT 25, 2018 | 9:00 AM
      Advances in Hydrogen-Deuterium Exchange Mass Spectrometry That Can Improve Studies of Biosimilars and Membrane Protein Drug Targets
      Advances in Hydrogen-Deuterium Exchange Mass Spectrometry That...
      Hydrogen-deuterium exchange mass spectrometry (HDX-MS) has developed into a powerful tool for investigating biopharmaceuticals, protein interactions, and membrane protein dynamics. Since 2010...
      Speaker: Jeffrey Hudgens, PhD
      Presented at: Analytical Chemistry Virtual Event Series 2018
      2
    NOV 21, 2019 | 8:00 AM
    How to maximize tumor microenvironment analysis from limited patient samples
    How to maximize tumor microenvironment analysis from limited patient...
    DATE: November 21, 2019TIME: 8:00am PSTA complex interaction among heterogenous immune and tumor cells within the tumorous tissue not only influences tumor progression, but can also sha...
    Speaker: César Evaristo, PhD
    Sponsored By: Miltenyi Biotec
    5 2
    NOV 18, 2019 | 7:00 AM
    Protect your cells with proper pipetting - How liquid handling influences your cell culture work
    Protect your cells with proper pipetting - How liquid handling...
    DATE: November 18, 2019TIME: 7:00am PST, 11:00am EST, 4:00pm CEWT How often do you pipette in your cell culture lab every day? Usually, we do it so often that we tend stop th...
    Speaker: Nadine Mellies, PhD , Katja Baesler, PhD
    Sponsored By: Eppendorf
    22 2
    NOV 07, 2019 | 10:00 AM
    Akoya Biosciences Webinar Series: Validating Multiplexed Antibody Panels for Pancreatic Research Applications
    Akoya Biosciences Webinar Series: Validating Multiplexed Antibody...
    DATE: November 7, 2019TIME: 10:00am PST, 1:00pm EST Studying the pathogenesis of diabetes requires detailed analysis of the pancreatic islet microenvironment and its numerous c...
    Speaker: Diane Saunders, PhD , Grady Carlson, PhD
    Sponsored By: Akoya Biosciences
    5
    OCT 31, 2019 | 9:00 AM
    Investigation of Endothelial Cell Viability and Growth on 3D printed GelMa Vascular Networks
    Investigation of Endothelial Cell Viability and Growth on 3D printed...
    DATE: October 31, 2019TIME: 9:00am PDT, 12:00pm EDT INTRODUCTION: A major limitation for the development of 3D engineered tissues is the absence of viable and perfusable...
    Speaker: Chrisoula Chappell
    Sponsored By: Thermo Fisher Scientific/Gibco
    8
    OCT 24, 2019 | 10:30 AM
    Supercritical fluid chromatography of polar biomolecules at the analytical and preparative scale: evaluation of global conformational changes in peptides and proteins
    Supercritical fluid chromatography of polar biomolecules at the...
    Supercritical fluid chromatography (SFC) has historically been very effective for exacting difficult chiral separations, with an unmatched power in terms of enabling highly efficient enantios...
    Speaker: Raffeal Bennett, PhD
    Presented at: Analytical Chemistry Virtual Event Series 2019
    2
     
     
  • OCT 24, 2019 | 6:00 AM
    Multidimension Chromatography; a Disruptive Technology - A Netflix Effect in the World of Chromatography!
    Multidimension Chromatography; a Disruptive Technology - A Netflix...
    Chromatography is the first choice in separation technique for a wide selection of applications, across many fields, ranging from simple to highly complex extracts. The main rationale for it...
    Speaker: Claude Mallet, PhD
    Presented at: Analytical Chemistry Virtual Event Series 2019
    2
    OCT 22, 2019 | 10:00 AM
    Demystifying Analytical Validation While Onboarding NGS Tests
    Demystifying Analytical Validation While Onboarding NGS Tests
    DATE: October 22, 2019TIME: 10:00am PDTNext-generation sequencing (NGS) is quickly becoming the go-to tool for testing labs looking to bring on additional clinically relevant tests requ...
    Speaker: Geoffrey Bien , Leah Ames, MS, MLS (ASCP) SCYM
    Sponsored By: Thermo Fisher Scientific
    5 3
    SEP 24, 2019 | 8:00 AM
    Interrogating 3D spheroid versus 2D monolayer cell models in drug discovery using cell health microplate assays
    Interrogating 3D spheroid versus 2D monolayer cell models in drug...
    DATE: September 24, 2019TIME: 7:00am PDTHigh throughput screening (HTS) is an effective method for identifying putative active compounds for therapeutics. Assays that evaluate changes i...
    Speaker: Dr. Adyary Fallarero, Ph.D. Adjunct Professor/Docent in Pharmaceutical Biology , Dr. Leticia Montoya, Ph.D., Chemistry and Biochemistry
    Sponsored By: Thermo Fisher Scientific
    9
    AUG 27, 2019 | 9:00 AM
    Akoya Biosciences Webinar Series:  Comprehensive Meta-analysis of Biomarker Technologies for Predictive Response of PD-1/PD-L1 Checkpoint Therapies
    Akoya Biosciences Webinar Series: Comprehensive Meta-analysis...
    DATE: August 27, 2019 TIME: 9:00am PDT, 12:00pm EDT Immunotherapies targeting PD-1 or PD-L1 have proven remarkably effective for treating cancer in some patients, with considerabl...
    Speaker: Cliff Hoyt, PhD , Steve Lu
    Sponsored By: Akoya Biosciences
    5
    JUL 24, 2019 | 8:00 AM
    Establishing a robust 37-marker mass cytometry assay for deep single-cell immune profiling of whole blood
    Establishing a robust 37-marker mass cytometry assay for deep...
    DATE: July 24, 2019TIMEL 8:00am PT Mass cytometry (CyTOF®) is a powerful approach to characterize the immune composition of complex biological specimens and gain insights in...
    Speaker: Adeeb Rahman, PhD , Mona Al-Maarri, PhD
    Sponsored By: Miltenyi Biotec
    8
     
     
  • JUN 05, 2019 | 5:00 PM
    Everyday culture practice - Improving reproducibility in cell culture
    Everyday culture practice - Improving reproducibility in cell...
    DATE: June 5, 2019TIME: 8:00am PDT, 11:00am EDT, 5:00pm CEST Eukaryotic cell cultures respond to the most subtle influence. Apart from the risk of contamination, minimal chan...
    Speaker: Jessica Wagener, PhD
    Sponsored By: Eppendorf
    23 2
    MAY 16, 2019 | 12:00 PM
    Isolation and expansion of mesenchymal stromal or stem cells (MSCs) for translational and clinical research
    Isolation and expansion of mesenchymal stromal or stem cells (MSCs)...
    DATE: May 16, 2019TIME: 12:00 PM PDT Mesenchymal stem or stromal cells (MSCs) are a promising resource for cell-based therapies and are being tested in already registered phase...
    Speaker: Kathrin Godthardt, B.Sc.
    Sponsored By: Miltenyi Biotec
    11
    MAY 16, 2019 | 4:00 PM
    Towards Standardized Workflows - A focus on microbiome insights
    Towards Standardized Workflows - A focus on microbiome insights
    DATE: May 16, 2019TIME: 7:00am PDT, 10:00am EDT, 4:00pm CEST The emergence of NGS is revolutionizing the microbiological sciences and transforming medicine. Deep sequencing has...
    Speaker: Rita Colwell, PhD , Christiane Honisch, PhD
    Sponsored By: Eppendorf
    10 2
    MAR 05, 2019 | 10:00 AM
    Comprehensive Capture of Human Neuropathology by Multiplexed Ion Beam Imaging (MIBI)
    Comprehensive Capture of Human Neuropathology by Multiplexed Ion...
    DATE: March 5, 2019TIME:  10:00am PSTSingle cell analysis, starting with the earliest low parameter fluorescent experiments, helped define the major cell subsets of human ce...
    Speaker: Sean Bendall, PhD
    Sponsored By: IONpath
    2 1
    DEC 05, 2018 | 8:00 PM
    The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary Tumors and in Circulation
    The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary...
    DATE: December 05, 2018TIME:  08:00 PSTWhat is desperately needed are methods to detect cells that cause metastasis–known as cancer stem cells (CSC), a subpopulation of...
    Speaker: Bruno Sainz, PhD
    Sponsored By: Thermo Fisher Scientific
    5 3
     
     
  • DEC 05, 2018 | 7:00 AM
    The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary Tumors and in Circulation
    The Use of Flow Cytometry to Identify Cancer Stem Cells in Primary...
    DATE: December 05, 2018TIME:  07:00 PSTWhat is desperately needed are methods to detect cells that cause metastasis–known as cancer stem cells (CSC), a subpopulation of...
    Speaker: Bruno Sainz, PhD
    Sponsored By: Thermo Fisher Scientific
    7
    NOV 29, 2018 | 9:00 AM
    An overview of primary immune deficiency disorders (PID) and laboratory testing for PID
    An overview of primary immune deficiency disorders (PID) and laboratory...
    DATE: November 29, 2018TIME:  9:00AM PDTAn overview of primary immune deficiency disorders (PID) and laboratory testing for PIDPrimary immune deficiencies (PIDs), oth...
    Speaker: Vijaya Knight, MD, PhD, (D)ABMLI
    Sponsored By: Binding Site
    10
    NOV 15, 2018 | 10:00 AM
    Multiplex fluorescence immunohistochemistry using the Ultivue InSituPlex platform on the Leica Biosystems BOND RX
    Multiplex fluorescence immunohistochemistry using the Ultivue...
    DATE:  November 15, 2018TIME:  10:00am PT, 1:00pm ET Multiplex fluorescence immunohistochemistry offers a window into the biology of human disease, enabling the ana...
    Speaker: Traci DeGeer, BS, HT (ASCP) HTL, QIHC , Alexander Klimowicz
    Sponsored By: Leica Biosystems
    6 1
    NOV 07, 2018 | 8:00 PM
    Analysis of Surface Antigens on Exosomes
    Analysis of Surface Antigens on Exosomes
    DATE: November 7, 2018TIME: 8:00PM PSTExosomes have been shown to have significant roles in cancer including disease progression acting in the tumor micro-environment, metasta...
    Speaker: Steve McClellan BS, MT, SCYM (ASCP)CM
    Sponsored By: Thermo Fisher Scientific
    3 1
    NOV 07, 2018 | 7:00 AM
    Analysis of Surface Antigens on Exosomes
    Analysis of Surface Antigens on Exosomes
    DATE: November 7, 2018TIME: 7:00AM PDTExosomes have been shown to have significant roles in cancer including disease progression acting in the tumor micro-environment, metastasis a...
    Speaker: Steve McClellan BS, MT, SCYM (ASCP)CM
    Sponsored By: Thermo Fisher Scientific
    11 1
     
     
  • NOV 01, 2018 | 12:00 AM
    Real-time Live-cell Analysis for Immunologists
    Real-time Live-cell Analysis for Immunologists
    Deriving deeper biological insight & improving productivity in immune-cell biologyThe most common in vitro methods for immunologists to analyze cells of the immune system are flow cyt...
    Speaker: Del Trezise, PhD
    Presented at: Dimension of Time: From Culture to Assay
    2
    OCT 25, 2018 | 12:00 PM
    Through the Looking Glass, and What Amino Acids Found There
    Through the Looking Glass, and What Amino Acids Found There
    Analytical chemistry can take you to strange places - for me it has been to the bottom of a quarry, being covered in dirt, excavating sesame-seed sized fossils that allow us to work out when...
    Speaker: Kirsty Penkman, PhD
    Presented at: Analytical Chemistry Virtual Event Series 2018
    1
    OCT 25, 2018 | 10:30 AM
    Trash or Treasure: Optimizing Titration Electrodes and Consumables
    Trash or Treasure: Optimizing Titration Electrodes and Consumables
    Everyone that uses titration in their lab knows how simple and fast the technique can be.  However, uncertainty around when to replace electrodes creates confusion. Reagent quality, tubi...
    Speaker: Jessica McVay , Lori Spafford
    Presented at: Analytical Chemistry Virtual Event Series 2018
    Sponsored By: Metrohm USA
    1
    OCT 25, 2018 | 9:00 AM
    Advances in Hydrogen-Deuterium Exchange Mass Spectrometry That Can Improve Studies of Biosimilars and Membrane Protein Drug Targets
    Advances in Hydrogen-Deuterium Exchange Mass Spectrometry That...
    Hydrogen-deuterium exchange mass spectrometry (HDX-MS) has developed into a powerful tool for investigating biopharmaceuticals, protein interactions, and membrane protein dynamics. Since 2010...
    Speaker: Jeffrey Hudgens, PhD
    Presented at: Analytical Chemistry Virtual Event Series 2018
    2
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