For evaluation of atherosclerotic processes, imaging of 3D structures in intact vascular tissues and functional aspects of the diseased artery are required. Multiphoton imaging has been applied in atherosclerosis to image murine vascular structures. However, homing of cells to the vessel wall has not been addressed. In our laboratory, we have established multiphoton scanning microscopy to allow 3D imaging of the entire structure of an isolated intact ApoE -/- mouse carotid artery, identifying homed fluorescently tagged adoptively transferred lymphocytes, in autofluorescent adventitial connective tissue and medial elastin fibres. Cell suspensions were prepared from peripheral and mesenteric lymph nodes of female C57BL/6 mice and labelled with Cell TrackerTM Red CMTPX (Molecular Probes Inc.). Lymphocytes (2-5 x 106) were injected i.v. into female ApoE -/- or C57BL/6 recipients. Twenty four hours after transfer excised intact mouse carotid arteries were imaged by two-photon microscopy. The use of this system provided us to clearly image lymphocyte infiltration mainly within the adventitia of an intact ApoE -/- mouse carotid artery. Consequently, this system could be a powerful tool to study immune cells behaviour in artery disease.
Life Sciences 2007 (2007) Proc Life Sciences, PC100
Poster Communications: Two-photon microscopy for imaging of lymphocytes recruitment in intact mouse carotid artery
P. Maffia1, 5, B. H. Zinselmeyer2, A. Ialenti1, S. Kennedy3, A. H. Baker4, I. B. McInnes5, J. M. Brewer2, P. Garside2
1. Department of Experimental Pharmacology, University of Naples Federico II, Naples, Italy. 2. Centre for Biophotonics , University of Strathclyde, Glasgow, United Kingdom. 3. Department of Physiology and Pharmacology, University of Strathclyde, Glasgow, United Kingdom. 4. BHF Glasgow Cardiovascular Research Centre , University of Glasgow, Glasgow, United Kingdom. 5. Division of Immunology, Infection & Inflammation, University of Glasgow, Glasgow, United Kingdom.
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