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In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis
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Lanthanide nanoparticles for high sensitivity multiparameter single cell analysis.

Jothirmayanantham Pichaandi1, Guangyao Zhao1, Alexandre Bouzekri2

  • 1Department of Chemistry , University of Toronto , 80 St George Street , Toronto , Ontario M5S 3H6 , Canada . Email: mnitz@chem.utoronto.ca ;

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|April 19, 2019
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Summary
This summary is machine-generated.

New nanoparticle (NP)-based reagents significantly enhance signal detection in mass cytometry (MC) for low-abundance biomarkers. These NP-antibody conjugates improve sensitivity for rare cell detection, complementing existing metal chelating polymer (MCP) reagents.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Immunology

Background:

  • Mass cytometry (MC) is a powerful tool for high-throughput, multiparameter biomarker analysis.
  • Current MC methods utilize antibodies conjugated to metal chelating polymers (MCPs) for mass tagging.
  • Improving sensitivity for low-abundance biomarkers remains a key challenge in MC.

Purpose of the Study:

  • To develop and evaluate nanoparticle (NP)-based reagents as novel mass tags for enhancing MC sensitivity.
  • To compare the performance of NP-antibody conjugates against traditional MCP-antibody conjugates.
  • To assess the utility of NP-based reagents for detecting rare cellular markers and cell types.

Main Methods:

  • Synthesis of silica-coated NaHoF4 NPs conjugated to antibodies.
  • Comparison of NP-antibody and MCP-antibody conjugate sensitivity across six cell lines and seven biomarkers.
  • Multi-parameter assay on peripheral blood mononuclear cells (PBMCs) using a cocktail of NP- and MCP-based reagents.

Main Results:

  • NP-antibody conjugates demonstrated significant signal enhancement for low-abundance biomarkers, particularly in direct detection methods (30-450 fold increase).
  • NP-reagents showed minimal non-specific binding and did not interfere with MCP-reagent function.
  • Rare CD14+ monocytes were identified with a 20-fold signal increase using NP-antibody conjugates compared to MCP-only methods.

Conclusions:

  • Nanoparticle-based reagents offer substantial signal amplification for mass cytometry, especially for detecting rare cellular targets.
  • NP-antibody conjugates represent a promising advancement for MC, expanding its capability to identify low-abundance biomarkers.
  • The combined use of NP- and MCP-based reagents provides a versatile approach for comprehensive cellular analysis.