Protocol for assessing and visualizing cell microaggregate formation in whole blood by imaging flow cytometry.

Area of Science:

• Hematology
• Biomedical Engineering
• Analytical Chemistry

Background:

• Blood cell aggregates serve as critical biomarkers in numerous medical conditions.
• Accurate quantification of these aggregates is essential for clinical diagnosis and monitoring.
• Existing methods may face challenges with sample preparation and artifact introduction.

Purpose of the Study:

• To develop and validate a protocol for accurate and quantitative analysis of blood cell aggregates.
• To utilize imaging flow cytometry for high-throughput analysis of cell aggregates in whole blood.
• To mitigate sample preparation artifacts and cell activation/morphological changes.

Main Methods:

• The protocol involves sample collection, staining, and measurement using imaging flow cytometry.
• Detailed gating strategies and cell morphology analysis are described.
• Erythrocyte lysis and leukocyte isolation were omitted to preserve sample integrity.

Main Results:

• The method allows for accurate and quantitative analysis of cell aggregates from small whole blood volumes.
• High-throughput imaging of leukocytes and platelets is achieved.
• Sample preparation artifacts, activation, and morphological changes are effectively mitigated.

Conclusions:

• This protocol offers a robust and efficient approach for analyzing blood cell aggregates.
• The method enhances the clinical utility of cell aggregates as biomarkers.
• It provides a reliable tool for research and diagnostics in hematology and related fields.