Immunothrombosis is a critical aspect affecting patients in acute care settings, especially in conditions involving infection, trauma, or severe inflammation. The interplay between hemostasis, innate immunity, and inflammation becomes highly relevant in these scenarios and can significantly impact patient outcomes. Blood cell aggregates are potential functional cellular biomarkers for prognostic and predictive biomarkers of immunothrombosis but require, due to the low logistical stability of the aggregates, a point-of-care (POC) solution for standardizable diagnostics. Here, we present a POC-compatible method that combines a quantitative phase imaging method with a microfluidic chip and a customized image analysis, resulting in a label- and sample preparation-free high-throughput imaging flow cytometer. We optimized our conditions to mimic the flow conditions in vessels at a low shear stress of ~1.000 s−1. In the acute care uses case, we demonstrate the dynamics of blood cell aggregates in critical care patients and discuss the potential impact on clinical workflows.
We discuss label-free high-throughput phase imaging, allowing the identification of platelet-platelet microaggregates and their components at single-platelet resolution in COVID-19 patients. The sample preparation-free measurement can be performed in 2 minutes with a benchtop-sized flow cytometer from a diluted blood sample matching requirement for point-of-care-testing in the ICU. Most importantly, the microfluidic solution allows for low shear stress to minimize loss of microaggregates yet achieving high throughput. We validated a positive correlation of (1) increased platelet-platelet concentration, (2) composition, and (3) size distribution with severity of COVID-19. Most importantly, the microaggregates we observe are in fact in a size regime of a single leukocyte, which allows for a sufficiently long circulation time in the peripheral blood system for quantification.
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