Expansion microscopy allows quantitative characterization of structural organization of platelet aggregates

Area of Science:

• Biophysics
• Cell Biology
• Medical Imaging

Background:

• Platelet aggregate structure is crucial for hemostasis and thrombosis, but current microscopy methods lack detailed volumetric imaging capabilities.
• Optical opacity and small platelet size hinder detailed analysis of platelet aggregate morphology.

Purpose of the Study:

• To develop and apply superresolution microscopy and image analysis techniques for detailed morphological description of platelet aggregates.
• To overcome limitations of current fluorescence microscopy in visualizing platelet aggregate structure.

Main Methods:

• Expansion microscopy was employed to enhance structural details of platelet aggregates.
• Custom image analysis workflows were developed for quantitative extraction of structural information.

Main Results:

• Expansion microscopy provided nanoscale resolution volumetric images of over 700 platelet aggregates.
• Quantitative analysis revealed platelet number, volume, morphology, and F-actin organization within aggregates.
• Structural disruptions caused by actin inhibitors (cytochalasin D, latrunculin A) were robustly detected.

Conclusions:

• A comprehensive workflow was established for qualitative and quantitative analysis of platelet aggregate architecture at multiple scales.
• This approach serves as a valuable tool for studying platelet aggregates and thrombi in various diseases, including cardiovascular disease and immunothrombosis.