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Atoms — and the protons, neutrons, and electrons that compose them — are extremely small. For example, a carbon atom weighs less than 2 × 10−23 g. When describing the properties of tiny objects such as atoms, we use appropriately small units of measure, such as the atomic mass unit (amu). The amu was originally defined based on hydrogen, the lightest element, then later in terms of oxygen. Since 1961, it has been defined with regard to the most abundant isotope of carbon, atoms of which...
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Sample Preparation for Mass Cytometry Analysis
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Ratiometric Barcoding for Mass Cytometry.

Xu Wu1,2,3, Quinn DeGottardi4, I-Che Wu1

  • 1Department of Chemistry , University of Washington , Seattle , Washington 98195 , United States.

Analytical Chemistry
|August 25, 2018
PubMed
Summary
This summary is machine-generated.

A new ratiometric lanthanide-coordinated polymer dots (Ln-Pdots) barcoding strategy enables 16 unique codes for mass cytometry. This method enhances high-throughput cellular analysis by eliminating labeling bias and improving sample resolution.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Materials Science

Background:

  • High-throughput cellular and molecular analysis relies on effective barcoding.
  • Existing barcoding methods may face challenges with reagent concentration variability and labeling efficiency.

Purpose of the Study:

  • To develop a novel ratiometric barcoding strategy for mass cytometry using lanthanide-coordinated polymer dots (Ln-Pdots).
  • To enhance the accuracy and throughput of cellular sample analysis.

Main Methods:

  • Developed ratiometric Ln-Pdots capable of generating 16 unique barcodes through combinations of 3 metal isotopes and 4 ratio intensity levels.
  • Applied the barcoding strategy to MCF-7 cells and peripheral blood mononuclear cells (PBMCs) using CD45-specific targeting.
  • Analyzed barcoded samples using mass cytometry.

Main Results:

  • Successfully generated and decoded 16 distinct barcodes for cell samples in mass cytometry.
  • Demonstrated high-mass-signal intensities and elimination of bias from reagent concentration and labeling efficiency.
  • Clearly distinguished 16 sets of labeled MCF-7 cells, showcasing excellent resolving power.
  • Validated the utility of ratiometric Ln-Pdot barcodes in multiplexed analysis of PBMCs.

Conclusions:

  • Ratiometric Ln-Pdot barcodes offer a robust solution for high-throughput mass cytometry.
  • The developed strategy improves accuracy and reduces variability in cellular analysis.
  • This technique is particularly valuable for precious human samples and multiplexed analyses.