Research Papers: Imaging

Quantification of volume, mass, and density of thrombus formation using brightfield and differential interference contrast microscopy

[+] Author Affiliations
Sandra M. Baker-Groberg, Kevin G. Phillips, Owen J. T. McCarty

Oregon Health and Science University, School of Medicine, Department of Biomedical Engineering, 3303 SW Bond Avenue, Portland, Oregon 97239

J. Biomed. Opt. 18(1), 016014 (Jan 25, 2013). doi:10.1117/1.JBO.18.1.016014
History: Received November 5, 2012; Revised December 6, 2012; Accepted December 7, 2012
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Abstract.  Flow chamber assays, in which blood is perfused over surfaces of immobilized extracellular matrix proteins, are used to investigate the formation of platelet thrombi and aggregates under shear flow conditions. Elucidating the dynamic response of thrombi/aggregate formation to different coagulation pathway perturbations in vitro has been used to develop an understanding of normal and pathological cardiovascular states. Current microscopy techniques, such as differential interference contrast (DIC) or fluorescent confocal imaging, respectively, do not provide a simple, quantitative understanding of the basic physical features (volume, mass, and density) of platelet thrombi/aggregate structures. The use of two label-free imaging techniques applied, for the first time, to platelet aggregate and thrombus formation are introduced: noninterferometric quantitative phase microscopy, to determine mass, and Hilbert transform DIC microscopy, to perform volume measurements. Together these techniques enable a quantitative biophysical characterization of platelet aggregates and thrombi formed on three surfaces: fibrillar collagen, fibrillar collagen +0.1nM tissue factor (TF), and fibrillar collagen +1nM TF. It is demonstrated that label-free imaging techniques provide quantitative insight into the mechanisms by which thrombi and aggregates are formed in response to exposure to different combinations of procoagulant agonists under shear flow.

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© 2013 Society of Photo-Optical Instrumentation Engineers

Citation

Sandra M. Baker-Groberg ; Kevin G. Phillips and Owen J. T. McCarty
"Quantification of volume, mass, and density of thrombus formation using brightfield and differential interference contrast microscopy", J. Biomed. Opt. 18(1), 016014 (Jan 25, 2013). ; http://dx.doi.org/10.1117/1.JBO.18.1.016014


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