This master’s thesis presents the methods we wanted to use for detection three elements of a blood clot - nanoparticles, fibrin and, red blood cells. Nanoparticles are organic or inorganic particles measuring one dimension less than 100 nm. Since their use recently increased, there are more mainly epidemiological studies linking nanoparticles to a variety of diseases, including vascular. One of the disease states
is the formation of a blood clot. A blood clot consists of fibrin, which polymerizes in the presence of blood cells - platelets and erythrocytes. The main question is whether inhaled nanoparticles present in polluted air can cause cardiovascular disease, or more specifically, the formation of a blood clot. The problem in studying possible causal link between nanoparticles and blood clot formation is the detection of all three components in a blood clot and the detection of nanoparticles whose size is below the resolution of classical fluorescence microscopy, so we wanted to find
a method to separate nanoparticles, fibrin, erythrocytes and platelets in the blood clot.
The methods we wanted to test for detection were confocal fluorescence microscopy along with stimulated emission depletion microscopy and fluorescence lifetime imaging
microscopy. All of the methods are based on fluorescence, so fluorescently labeled titanium dioxide nanotubes and fluorescently labeled fibrin were used. In red blood
cells, we detected auto-fluorescence of hemoglobin.
We developed a method by which we were able to separate both cell types (erythrocytes and platelets), fibrin and nanoparticles. The method used should be further improved in the future so that they could be used not only for fixed samples but also in real-time experiments.
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