Development of a fluorescent method for detecting the onset of coagulation in human plasma on microstructured lateral flow platforms

Abstract

Microfluidic devices and microsystems have been used to develop blood coagulation monitoring devices for point of care diagnostic use. However, many of them suffer from inherent variability and imprecision, partly due to the fact that they only detect changes in bulk clotting properties and do not reflect the microscopic nature of blood coagulation. This work demonstrates microstructured lateral flow platforms used in combination with fluorescently labelled fibrinogen to detect microscopic clot formation. Plasma samples applied to platforms modified with coagulation activation reagents and fluorescent fibrinogen produced changes in fluorescence intensity due to incorporation of the fluorophore into the forming microclots. It was found that the change in the distribution of the fluorescence within the sample over time was an excellent predictor of the onset of coagulation, which could be used to determine the clotting time. The impact of various assay parameters was optimised and the assay was shown to be capable of measuring the effect of heparin concentration on blood clotting time from 0 to 1.5 U mL⁻¹.