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Method Article
Johan Heemskerk
Heemskerk lab, Department of Biochemistry, Maastricht University
Corresponding Author
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Thrombus formation by adhering and aggregating blood platelets is fundamental to hemostasis and is a prerequisite for vascular occlusion in pathological thrombosis. The parallel-plate flow chamber technique has been extensively used to measure platelet adhesion and activation in vitro at arterial or venous flow conditions. Here, we describe the use of brightfield and confocal fluorescence microscopy to record the various platelet activation processes contributing to thrombus formation on microspotted arrays of thrombogenic surfaces; and we give procedures to analyze the acquired microscopic images. Furthermore, we describe technical problems that can be expected using the microspot technique. Content: (A) Flow chamber preparation and whole blood perfusion. (B) Brightfield and fluorescence microscopic imaging of thrombi. (C) Analysis of brightfield and fluorescence images.
Keywords
Thrombus formation, platelets, multispot technique, multiparameter analysis,
Figure 1
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Associated Publications
Identification of platelet function defects by multi-parameter assessment of thrombus formation
by Susanne M. de Witt, Frauke Swieringa, Rachel Cavill, +12
Nature Communications (17 July, 2014)
Method Article
Johan Heemskerk
Heemskerk lab, Department of Biochemistry, Maastricht University
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 26 Aug, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Thrombus formation by adhering and aggregating blood platelets is fundamental to hemostasis and is a prerequisite for vascular occlusion in pathological thrombosis. The parallel-plate flow chamber technique has been extensively used to measure platelet adhesion and activation in vitro at arterial or venous flow conditions. Here, we describe the use of brightfield and confocal fluorescence microscopy to record the various platelet activation processes contributing to thrombus formation on microspotted arrays of thrombogenic surfaces; and we give procedures to analyze the acquired microscopic images. Furthermore, we describe technical problems that can be expected using the microspot technique. Content: (A) Flow chamber preparation and whole blood perfusion. (B) Brightfield and fluorescence microscopic imaging of thrombi. (C) Analysis of brightfield and fluorescence images.
Figure 1
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Associated Publications
Identification of platelet function defects by multi-parameter assessment of thrombus formation
by Susanne M. de Witt, Frauke Swieringa, Rachel Cavill, +12
Nature Communications (17 July, 2014)