High correlation between 2 flow cytometry platforms in the microparticles analysis using a new calibrated beads...

Area of Science:

• Biomedical Science
• Clinical Chemistry
• Flow Cytometry

Background:

• Microparticles (MPs) are valuable noninvasive biomarkers for diagnosing and prognosing various pathologic conditions.
• Significant variability in MP results stems from a lack of standardized preanalytical and analytical methods.
• Megamix-Plus beads offer a potential solution for standardizing MP analysis on specific flow cytometer platforms.

Purpose of the Study:

• To evaluate the comparability of total MPs and platelet-derived MPs (PMPs) counts between two different flow cytometer platforms.
• To assess the effectiveness of Megamix-Plus beads in standardizing MP measurements across diverse cytometer systems.

Main Methods:

• Blood samples from 65 deep venous thrombosis patients were processed to obtain platelet-poor plasma (PPP).
• Total MPs and PMPs were quantified using two flow cytometers: LSRFortessa (Becton Dickinson) and Navios (Beckman Coulter).
• Megamix-Plus beads were used for calibration, with side scatter beads for LSRFortessa and forward scatter beads for Navios.

Main Results:

• A high correlation was observed for both total MPs (r = 0.908, P < 0.01) and PMPs (r = 0.910, P < 0.001) between the two platforms.
• Absolute counts of total MPs and PMPs were significantly higher when measured using the Navios flow cytometer compared to the LSRFortessa.
• While both platforms are viable for MP determination, differences in absolute counts necessitate careful consideration.

Conclusions:

• Megamix-Plus beads facilitate standardization, showing high correlation in MP and PMP measurements across different flow cytometers.
• Despite high correlation, absolute MP and PMP counts vary significantly between platforms, impacting direct comparison.
• For multicenter studies, using identical platforms and calibration tools is recommended for consistent MP analysis.