Cutting-edge analysis of extracellular microparticles using ImageStream(X) imaging flow cytometry

Area of Science:

• Extracellular vesicle biology
• Biomedical engineering
• Cellular biology

Background:

• Extracellular vesicles (EVs) play vital roles in intercellular communication and pathophysiology.
• Accurate quantification of EVs is essential for their use as preclinical and clinical tools.
• Traditional flow cytometry methods lack sensitivity and accuracy for EV analysis.

Purpose of the Study:

• To evaluate the Amnis ImageStream(X) Mk II imaging flow cytometer for accurate microparticle quantification.
• To assess the utility of imaging flow cytometry for EV analysis in complex biological samples.

Main Methods:

• Utilized Amnis ImageStream(X) Mk II imaging flow cytometer for microparticle analysis.
• Analyzed calibration beads (1 µm to 20 nm) and microparticles in whole blood, plasma, and leukocyte supernatants.
• Employed minimal sample preparation and volume.

Main Results:

• The ImageStream(X) Mk II accurately analyzed microparticles across a size range of 20 nm to 1 µm.
• Quantification was successful in various sample types, including whole blood and plasma.
• Simultaneous phenotypic definition of parent cells and offspring microparticles was achieved.
• Real-time microparticle generation kinetics were monitored.

Conclusions:

• The Amnis ImageStream(X) Mk II provides accurate and sensitive analysis of microparticles.
• This imaging flow cytometry approach overcomes limitations of traditional methods.
• The ImageStream(X) Mk II is a valuable tool for advancing extracellular vesicle research.