Quantitative evaluation of morphological changes in activated platelets in vitro using digital holographic microscopy

Area of Science:

• Biomedical Engineering
• Hematology
• Microscopy

Background:

• Conventional platelet aggregometry is limited for long-term quantitative analysis of platelet aggregation and morphology.
• Digital holographic microscopy (DHM) allows label-free, non-destructive quantitative evaluation of cell size and morphology.

Purpose of the Study:

• To validate the applicability of DHM for quantitatively assessing platelet activation and aggregation.
• To modify image analysis techniques for evaluating aggregated platelets.

Main Methods:

• Platelet-rich plasma was activated and subjected to DHM, SEM, AFM, optical microscopy, and FCM.
• Time-course data were analyzed by plotting average optical thickness versus attachment area.
• Aggregated platelets were identified and analyzed as single cells.

Main Results:

• DHM successfully quantified morphological changes in activated platelets.
• Platelet distribution shifted from a 'control' region to an 'activated' region with increasing activation time.
• DHM findings were consistent with SEM and FCM data.

Conclusions:

• DHM is a validated, promising tool for quantitative platelet function testing.
• DHM enables evaluation of activation-dependent platelet morphological changes.
• DHM has potential applications in platelet concentrate quality assurance, diagnostics, and drug discovery.