Abstract
A new biosensor for the real-time analysis of thrombus formation is reported. The fast and accurate monitoring of the individual thrombotic risk represents a challenge in cardiovascular diagnostics and in treatment of hemostatic diseases. Thrombus volume, as representative index of the related thrombotic status, is usually estimated with confocal microscope at the end of each in vitro experiment, without providing a useful behavioral information of the biological sample such as platelets adhesion and aggregation in flowing blood. Our device has been developed to work either independently or integrated with the microscopy system; thus, images of the fluorescently labeled platelets are acquired in real-time during the whole blood perfusion, while the global electrical impedance of the blood sample is simultaneously monitored between a pair of specifically designed gold microelectrodes. Fusing optical and electrical data with a novel technique, the dynamic of thrombus formation events in flowing blood can be reconstructed in real-time, allowing an accurate extrapolation of the three-dimensional shape and the spatial distribution of platelet thrombi forming and growing within artificial capillaries. This biosensor is accurate and it has been used to discriminate different hemostatic conditions and to identify weakening and detaching platelet aggregates. The results obtained appear compatible with those quantified with the traditional optical method. With advantages in terms of small size, user-friendliness and promptness of response, it is a promising device for the fast and automatic individual health monitoring at the Point of Care (POC).
Highlights
Blood coagulation is a complex mechanism whose alteration can produce diseases such as thrombosis, embolisms and hemorrhages
If the specific aim is to measure the volumetric structure of an object, confocal microscope provides as output a sequence of fluorescent images, called z-stack, acquired along the vertical z-dimension, that should facilitate the three-dimensional (3D) volume reconstruction and measurement, rejecting the out-of-focus fluorescence light
This paper further explores the relevance of blood impedance technique in measuring thrombus formation under flow conditions
Summary
Blood coagulation is a complex mechanism whose alteration can produce diseases such as thrombosis, embolisms and hemorrhages. The advances in technologies and computational methods have moved the cell biology toward a quantitative view [4, 5]: there is an increasing interest for an in vitro measurement of coagulation process [6, 7] in order to determine, as quickly as possible, the risk of thrombosis or bleeding, expecially during a surgical treatment. The improvements in microscope technologies and computational techniques make possible to investigate platelets functionality and thrombus formation [4, 15]. The accuracy and the standardization of thrombus volume measurements, starting from confocal microscope images, have always been open and problematic issues in the specific biological field of blood coagulation. The image acquisition and post process from confocal microscope is a costly and laborious methodology; for this reason its application in clinics is restricted
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