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OMIP-097: High-parameter phenotyping of human platelets by spectral flow cytometry.

Cytometry. Part A : the journal of the International Society for Analytical Cytology·2023
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Signal without noise: Practical antibody titration for platelet flow cytometry.

Benjamin E J Spurgeon1

  • 1Versiti, Milwaukee, Wisconsin, USA.

Cytometry. Part B, Clinical Cytometry
|December 19, 2025
PubMed
Summary

Optimizing antibody titration for platelet flow cytometry is crucial for accurate analysis. This guide offers practical strategies for whole blood assays to enhance signal resolution and assay reproducibility.

Keywords:
blood plateletsflow cytometryfluorescent dyesplatelet activationplatelet function testsresearch design

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Area of Science:

  • Immunology
  • Hematology
  • Analytical Chemistry

Background:

  • Platelet flow cytometry is vital for assessing platelet characteristics.
  • Standardized antibody titration protocols are needed for reliable results.

Purpose of the Study:

  • To provide practical methods for antibody titration in platelet flow cytometry.
  • To optimize staining for both constitutive and inducible platelet markers.

Main Methods:

  • Whole blood antibody titration strategies for various markers.
  • Use of controls, consideration of coexpression, and subsaturating concentrations.
  • Introduction of stain index and separation index for performance evaluation.

Main Results:

  • Defined quantitative metrics for assessing staining quality.
  • Addressed technical variables like combinatorial titration and spillover spreading.
  • Recommended titration under final assay conditions for biological relevance.

Conclusions:

  • Developed robust strategies for high-resolution platelet assays.
  • Enhanced reproducibility and biological relevance in flow cytometry.
  • Provided a foundation for automated and standardized platelet analysis.