High-Resolution Imaging of Intravascular Atherogenic Inflammation in Live Mice

Raphael Chevre, Jose Maria Gonzalez-Granado, Remco T. A. Megens, Vinatha Sreeramkumar, Carlos Silvestre-Roig, Pedro Molina-Sanchez, Christian Weber, Oliver Soehnlein, Andres Hidalgo*, Vicente Andres

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

55 Citations (Web of Science)


Rationale: The inflammatory processes that initiate and propagate atherosclerosis remain poorly understood, largely because defining the intravascular behavior of immune cells has been technically challenging. Respiratory and pulsatile movements have hampered in vivo visualization of leukocyte accumulation in athero-prone arteries at resolutions achieved in other tissues. Objective: To establish and to validate a method that allows high-resolution imaging of inflammatory leukocytes and platelets within the carotid artery of atherosusceptible mice in vivo. Methods and Results: We have devised a procedure to stabilize the mouse carotid artery mechanically without altering blood dynamics, which dramatically enhances temporal and spatial resolutions using high-speed intravital microscopy in multiple channels of fluorescence. By applying this methodology at different stages of disease progression in atherosusceptible mice, we first validated our approach by assessing the recruitment kinetics of various leukocyte subsets and platelets in athero-prone segments of the carotid artery. The high temporal and spatial resolution allowed the dissection of both the dynamic polarization of and the formation of subcellular domains within adhered leukocytes. We further demonstrate that the secondary capture of activated platelets on the plaque is predominantly mediated by neutrophils. Finally, we couple this procedure with triggered 2-photon microscopy to visualize the 3-dimensional movement of leukocytes in intimate contact with the arterial lumen. Conclusions: The improved imaging of diseased arteries at subcellular resolution presented here should help resolve many outstanding questions in atherosclerosis and other arterial disorders.
Original languageEnglish
Pages (from-to)770-779
JournalCirculation Research
Issue number5
Publication statusPublished - 28 Feb 2014


  • atherosclerosis
  • blood platelets
  • carotid arteries
  • neutrophils

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