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A CD45-based barcoding approach to multiplex mass-cytometry (CyTOF).

Liyun Lai1, Raymond Ong, Juntao Li

  • 1SingHealth Translational Immunology and Inflammation Centre, Singapore Health Services Pte Ltd, Singapore.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|February 4, 2015
PubMed
Summary
This summary is machine-generated.

This study introduces a new method for barcoding immune cells using CD45 antibodies and isotopes, improving mass cytometry (CyTOF) analysis. This technique enhances sensitivity and user-friendliness, especially for scarce samples.

Keywords:
CD45CyTOFbarcodingmultiplexing

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Area of Science:

  • Immunology
  • Analytical Chemistry
  • Biotechnology

Background:

  • Cytometry by time-of-flight (CyTOF) offers high-dimensional immune system analysis.
  • Limited sample availability can hinder CyTOF's full potential.
  • Existing maleimido-monoamide-DOTA (m-DOTA) barcoding methods face signal intensity challenges.

Purpose of the Study:

  • To develop an alternative barcoding strategy for CyTOF using antibody-isotope conjugates.
  • To overcome limitations of m-DOTA, particularly signal intensity and stability.
  • To enable effective CyTOF analysis with scarce or precious samples.

Main Methods:

  • Conjugating antibodies specific for the CD45 membrane protein with different isotopes.
  • Designing experiments to test various culture types and intracellular/extracellular targets.
  • Evaluating the new method against m-DOTA for sensitivity, specificity, and flexibility.

Main Results:

  • The CD45 antibody-isotope barcoding approach demonstrates high sensitivity and specificity.
  • The method proves flexible for analyzing both extracellular and intracellular targets.
  • This technique offers improved stability and user-friendliness compared to m-DOTA.

Conclusions:

  • Antibody-isotope conjugation targeting CD45 provides a robust alternative for immune cell barcoding in CyTOF.
  • This method effectively addresses the limitations of m-DOTA, enhancing data reliability.
  • The developed strategy expands the utility of CyTOF, particularly for studies with limited sample quantities, such as clinical samples.