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In vivo real-time multiphoton imaging of T lymphocytes in the mouse brain after experimental stroke

Fumagalli, Stefano and Coles, Jonathan A and Ejlerskov, Patrick and Ortolano, Fabrizio and Bushell, Trevor J and Brewer, James M and De Simoni, Maria-Grazia and Dever, Gary and Garside, Paul and Maffia, Pasquale and Carswell, Hilary V (2011) In vivo real-time multiphoton imaging of T lymphocytes in the mouse brain after experimental stroke. Stroke, 42 (5). pp. 1429-1436. ISSN 0039-2499

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Abstract

BACKGROUND AND PURPOSE: To gain a better understanding of T cell behavior after stroke, we have developed real-time in vivo brain imaging of T cells by multiphoton microscopy after middle cerebral artery occlusion. METHODS: Adult male hCD2-GFP transgenic mice that exhibit green fluorescent protein-labeled T cells underwent permanent left distal middle cerebral artery occlusion by electrocoagulation (n=6) or sham surgery (n=6) and then multiphoton laser imaging 72 hours later. RESULTS: Extravasated T cell number significantly increased after middle cerebral artery occlusion versus sham. Two T cell populations existed after middle cerebral artery occlusion, possibly driven by 2 T cell subpopulations; 1 had significantly lower and the other significantly higher track velocity and displacement rate than sham. CONCLUSIONS: The different motilities and behaviors of T cells observed using our imaging approach after stroke could reveal important mechanisms of immune surveillance for future therapeutic exploitations.