A NEW APPARATUS BY COMBINATION WITH A MODYFIED CONE-PLATE TYPE VISCOMETER AND EPIFLUORESCENCE MICROSCOPY TO EVALUATE ANTI-THROMBOGENIC BIOMATERIALS

Abstract

We developed a new apparatus to evaluate platelet-biomaterial interactions, the initial event of thrombus formation, in real-time with relatively small platelet suspension volumes under shear flow conditions. This is the first report on the observation of platelets adhered to opaque biomaterials in real-time by an apparatus consisting of a modified cone and plate-type viscometer combined with an upright epi-fluorescence microscope. Because no difference was observed between the number of labeled and non-labeled platelets with CFSE (a fluorescent dye) adhered to the materials, it was suggested that CFSE has no affect on platelet adhesion to test materials or platelet activation. Using this apparatus, platelets adhered to the ePTFE which is opaque and used as an artificial vessel under three types of shear flow were clearly visualized in real-time by epi-illuminated light. Our findings show that our cone and plate-type viscometer-based apparatus is a useful instrument for testing and screening biomaterials and provides a precise quantitative evaluation of platelet adhesion.