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J Biomed Opt. 2015 Feb;20(2):26005. doi: 10.1117/1.JBO.20.2.026005.

Intravital live cell triggered imaging system reveals monocyte patrolling and macrophage migration in atherosclerotic arteries.

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La Jolla Institute, 9420 Athena Circle, La Jolla, California 92037, United StatesbUniversity of California, Department of Bioengineering, San Diego, 9500 Gilman Drive, La Jolla, California 92093, United States.
La Jolla Institute, 9420 Athena Circle, La Jolla, California 92037, United StatescWroclaw Research Centre EIT+, Stablowicka 147, 54-066 Wroclaw, Poland.
University of Alberta, Department of Computing Science, 8900 114 Street Northwest, Edmonton, Alberta T6G 2S4, Canada.
La Jolla Institute, 9420 Athena Circle, La Jolla, California 92037, United States.


Intravital multiphoton imaging of arteries is technically challenging because the artery expands with every heartbeat, causing severe motion artifacts. To study leukocyte activity in atherosclerosis, we developed the intravital live cell triggered imaging system (ILTIS). This system implements cardiac triggered acquisition as well as frame selection and image registration algorithms to produce stable movies of myeloid cell movement in atherosclerotic arteries in live mice. To minimize tissue damage, no mechanical stabilization is used and the artery is allowed to expand freely. ILTIS performs multicolor high frame-rate two-dimensional imaging and full-thickness three-dimensional imaging of beating arteries in live mice. The external carotid artery and its branches (superior thyroid and ascending pharyngeal arteries) were developed as a surgically accessible and reliable model of atherosclerosis. We use ILTIS to demonstrate Cx3cr1GFP monocytes patrolling the lumen of atherosclerotic arteries. Additionally, we developed a new reporter mouse (Apoe−/−Cx3cr1GFP/+Cd11cYFP) to image GFP+ and GFP+YFP + macrophages “dancing on the spot” and YFP+ macrophages migrating within intimal plaque. ILTIS will be helpful to answer pertinent open questions in the field, including monocyte recruitment and transmigration, macrophage and dendritic cell activity, and motion of other immune cells.

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