Two-photon intravital imaging of thrombus development.
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.
Kamocka
Malgorzata M
MM
Indiana University School of Medicine, Department of Medical and Molecular Genetics, Indianapolis, Indiana 46202, USA. mkamocka@iupui.edu
Mu
Jian
J
Liu
Xiaomin
X
Chen
Nan
N
Zollman
Amy
A
Sturonas-Brown
Barbara
B
Dunn
Kenneth
K
Xu
Zhiliang
Z
Chen
Danny Z
DZ
Alber
Mark S
MS
Rosen
Elliot D
ED
D000465:Algorithms; D000818:Animals; D003196:Computer Graphics; D003198:Computer Simulation; D004195:Disease Models, Animal; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007834:Lasers; D008643:Mesentery / Q000293:injuries; D051379:Mice; D008810:Mice, Inbred C57BL; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D018715:Microscopy, Video; D013499:Surface Properties; D013927:Thrombosis / Q000378:metabolism / Q000473:pathology*