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. Author manuscript; available in PMC: 2017 Feb 1.
Published in final edited form as: Cytometry A. 2015 Feb 20;89(2):111–122. doi: 10.1002/cyto.a.22647

Table 3.

Overview of major challenges related to flow cytometric analysis of MPs.

Important Issues in MP Analysis by Flow Cytometry
Pre-Analytical Analytical/Technical

  • Method of blood collection

    • Tourniquet use

    • Needle diameter

    • Type of anticoagulant

  • Sample processing

    • Sample type (whole blood, plasma, isolated MPs)

    • Time to sample preparation

    • Centrifugation protocol

    • “Micro-clot” formation

  • Sample handling and storage

    • Sample transportation/agitation

    • Fresh vs freeze/thaw

  • Flow cytometer

    • Intrinsic resolution capabilities

  • Size gating

    • Beads vs biologicals

  • Fluorescence gating

    • Proper use of isotype controls (ITCs)

    • Detection of dimly expressed antigens

    • Titration of antibodies and ITCs

    • Fluorochrome aggregates

  • MP enumeration

    • Use of counting beads

    • “Swarm effect”