Abstract
IntroductionCurrent developments to assess qualitative and quantitative platelet traits in flowed whole-blood are based on microfluidic devices that mostly operate at room temperature. However, operation at physiological temperature (37 °C) may increase the assay's sensitivity, and facilitates the comparison to other platelet function tests of the diagnostic laboratory. Materials and methodsWe adapted the conventional microspot-based microfluidic device with a simple thermo-coupled pre-heating module. Automated analysis of microscopic images assisted in obtaining five time-dependent parameters of thrombus formation over collagen microspots (shear rate 1000 s−1). These modifications allowed rapid testing of control and patient blood samples at physiological temperature. Results and conclusionThe higher temperature enhanced platelet adhesion and aggregation as well as late thrombus characteristics such as size and contraction, when compared to room temperature. Moreover, assessment at 37 °C indicated a time-dependent impairment of the thrombus parameters in blood from patients taking common antiplatelet medication, i.e. aspirin and/or clopidogrel. This pointed to increased contribution of the autocrine platelet agonists thromboxane A2 and ADP in the buildup of contracted thrombi under flow. Overall, this study underlined the advantage of multiparameter assessment of microfluidic thrombus formation in detecting an acquired platelet dysfunction, when operating at physiological temperature. This work may bring microfluidics tests closer to the diagnostic laboratory.
Highlights
Current developments to assess qualitative and quantitative platelet traits in flowed whole-blood are based on microfluidic devices that mostly operate at room temperature
We used a microfluidic system (Fig. 1A–C), in which recalcified whole-blood was flowed over collagen microspots at room temperature (RT) and thrombus formation was assessed from micro scopic images in a multiparameter way, to characterize platelet-related abnormalities [18,19,20]
We adapted this system to operate at physiological temperature of 37 ◦C, in particular to better link the results to common platelet function tests performed in the diagnostic laboratory
Summary
Current developments to assess qualitative and quantitative platelet traits in flowed whole-blood are based on microfluidic devices that mostly operate at room temperature. Assess ment at 37 ◦C indicated a time-dependent impairment of the thrombus parameters in blood from patients taking common antiplatelet medication, i.e. aspirin and/or clopidogrel This pointed to increased contribution of the autocrine platelet agonists thromboxane A2 and ADP in the buildup of contracted thrombi under flow. Several point-of-care tests are available that directly assess platelet function in whole-blood, such as the platelet function analyzer (PFA), rotational thromboelastography (platelet modules) and whole blood aggregometry (Multiplate), all of which are operating at physiological temperature of 37 ◦C [1,7,10] Several of these are used in the peri-operative setting, a drawback is lack of sensitivity in part due to the absence of flow [11,12,13]
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