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Updated: Jun 15, 2026
Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli
Published on: September 25, 2016
Two-photon intravital imaging of thrombus development
Malgorzata M Kamocka1, Jian Mu, Xiaomin Liu
1Indiana University School of Medicine, Department of Medical and Molecular Genetics, Indianapolis, Indiana 46202, USA. mkamocka@iupui.edu
Abstract:
Thrombus development in mouse mesenteric vessels following laser-induced injury was monitored by high-resolution, near-real-time, two-photon, intravital microscopy. In addition to the use of fluorescently tagged fibrin(ogen) and platelets, plasma was labeled with fluorescently tagged dextran. Because blood cells exclude the dextran in the single plane, blood cells appear as black silhouettes. Thus, in addition to monitoring the accumulation of platelets and fibrin in the thrombus, the protocol detects the movement and incorporation of unlabeled cells in and around it. The developing thrombus perturbs the blood flow near the thrombus surface, which affects the incorporation of platelets and blood cells into the structure. The hemodynamic effects and incorporation of blood cells lead to the development of thrombi with heterogeneous domain structures. Additionally, image processing algorithms and simulations were used to quantify structural features of developing thrombi. This analysis suggests a novel mechanism to stop the growth of developing thrombus.
Insights
This study visualizes thrombus development in mice using advanced microscopy. It reveals how blood flow changes and cell incorporation create complex thrombus structures, suggesting a new way to halt their growth.
Area of Science:
- Vascular biology
- Biomedical imaging
- Hemodynamics
Background:
- Thrombus formation is a complex process involving platelets and fibrin.
- Understanding thrombus development is crucial for treating thrombotic diseases.
- Current methods have limitations in visualizing dynamic thrombus growth in vivo.
Purpose of the Study:
- To visualize and quantify thrombus development in mouse mesenteric vessels in near real-time.
- To investigate the role of hemodynamics and cell incorporation in thrombus structure.
- To identify novel mechanisms regulating thrombus growth.
Main Methods:
- High-resolution, two-photon intravital microscopy was employed.
- Fluorescently tagged fibrinogen, platelets, and plasma dextran were used for labeling.
- Image processing algorithms and simulations quantified structural features.
Main Results:
- Near real-time visualization of thrombus development was achieved.
- Blood flow perturbation near the thrombus surface was observed.
- Heterogeneous thrombus structures formed due to hemodynamic effects and cell incorporation.
Conclusions:
- Dynamic imaging revealed complex thrombus development influenced by flow and cell dynamics.
- A novel mechanism for inhibiting thrombus growth was suggested.
- This approach provides new insights into thrombus formation and potential therapeutic targets.
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