Detection of platelet-monocyte aggregates by the ADAM(®) image cytometer

Int J Med Sci. 2014 Sep 18;11(12):1228-33. doi: 10.7150/ijms.10008. eCollection 2014.

Abstract

Background: Inappropriate platelet activation is known to be associated with various thrombotic disorders. Platelet-monocyte aggregates (PMAs), whose formation is mediated by platelet surface P-selectin (CD62P), can be used as a reliable marker to detect platelet activation. Previous studies have generally detected PMAs through flow cytometry-based approaches. Recently, the ADAM(®) image cytometer (Nanoentek Inc., Seoul, Korea) was developed for image-based cellular analysis. In this study, we detected PMAs with the ADAM(®) cytometer, evaluated the reproducibility of the measurements made by the ADAM(®) cytometer, and compared the abilities of the ADAM(®) cytometer and a flow cytometric assay to detect PMAs.

Methods: Whole blood samples were collected from patients. Within 5 minutes of collection, anticoagulated whole blood samples were fixed in 10% paraformaldehyde and 5% glyoxal. Nineteen clinical specimens were collected; each was analyzed three times with the ADAM(®) cytometer in order to assess the reproducibility of its measurements. To compare the ability of the ADAM(®) cytometer with that of a flow cytometer to detect PMAs, each cytometer was used for 23 clinical samples and the correlation of the measurements was determined.

Results: The PMA measurements made by the ADAM(®) cytometer showed good reproducibility (CV < 10% for all specimens). Moreover, the PMA measurements made by the ADAM(®) cytometer exhibited a high correlation with those made by a flow cytometric assay (R = 0.944).

Conclusions: The ADAM(®) cytometer is a suitable alternative method to the flow cytometry-based assays. Since the ADAM cytometer does not need specialized instrument knowledge or software proficiency (unlike flow cytometry), the ADAM(®) cytometer can be used as a rapid and reliable POCT device to measure platelet activation in peripheral blood. This, in turn, will provide valuable information regarding patient propensities to thrombotic diseases.

Keywords: ADAM®; image cytometer; platelet activation; platelet-monocyte aggregates.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Algorithms
  • Blood Platelets / cytology*
  • Cell Aggregation
  • Child
  • Child, Preschool
  • Female
  • Flow Cytometry / instrumentation*
  • Flow Cytometry / methods
  • Flow Cytometry / statistics & numerical data
  • Fluorescent Dyes
  • Humans
  • Male
  • Middle Aged
  • Monocytes / cytology*
  • Platelet Activation*
  • Platelet Aggregation
  • Reproducibility of Results
  • Thrombosis / blood
  • Young Adult

Substances

  • Fluorescent Dyes