Measurement of in vitro P-selectin expression by flow cytometry

Sean D. Kennedy, Yasuyuki Igarashi, Thomas Stephen Kickler

Research output: Contribution to journalArticle

Abstract

Measurement of in vivo platelet activation is difficult after phlebotomy and during blood processing for analysis. We used flow cytometry to measure platelet surface expression of P-selectin in the presence and absence of trimethylsphingosine (a platelet activation inhibitor) and compared the results with those from the standard methods of preventing in vitro P- selectin expression. Percent activation was calculated as a ratio of mean sample fluorescence to 100% mean fluorescence after phorbol myristate acetate treatment. Twenty-five micromoles per liter of trimethylsphingosine kept in vitro platelet activation below 5% up to 6 hours after collection and below 10% at 24 hours after collection. Trimethylsphingosine failed to prevent platelet activation caused by centrifugation, storage at 4°C, or stimulation with common agonists. Addition of trimethylsphingosine to whole blood was valuable in preventing in vitro platelet activation. This compound promises to be a useful preservative for diagnostic testing of platelet activation.

Original languageEnglish (US)
Pages (from-to)99-104
Number of pages6
JournalAmerican Journal of Clinical Pathology
Volume107
Issue number1
StatePublished - Jan 1997

Fingerprint

P-Selectin
Platelet Activation
Flow Cytometry
Fluorescence
Phlebotomy
Platelet Aggregation Inhibitors
Tetradecanoylphorbol Acetate
Centrifugation
In Vitro Techniques
Blood Platelets

Keywords

  • Coagulation
  • Flow cytometry
  • Integrins
  • P-selectin
  • Platelet function

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Measurement of in vitro P-selectin expression by flow cytometry. / Kennedy, Sean D.; Igarashi, Yasuyuki; Kickler, Thomas Stephen.

In: American Journal of Clinical Pathology, Vol. 107, No. 1, 01.1997, p. 99-104.

Research output: Contribution to journalArticle

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