Affiliations: GKSS-Forschungszentrum Geesthacht, Berlin–Brandenburger Zentrum für Regenerative Therapien, Campus Virchow-Klinikum der Charité, Universitätsmedizin Berlin, Berlin, Germany | Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Freie Universität Berlin, Berlin, Germany | Central Institute for Biomedical Engineering, University of Ulm, Ulm, Germany | Department of Clinical Hemostasiology and Transfusion Medicine, University of Saarland, Homburg, Germany
Abstract: Several methods are available today for the investigation of microcirculation in animal models, but they can be invasive and time-consuming depending on the area investigated. In particular, non-invasive methods that can be conducted rapidly and without dye or tracer injections are in demand. The cutaneous microcirculation can be easily studied in the dorsal corium limbi of the third toe of the porcine forelimb using intravital microscopy – analogous to nail fold capillary microscopy in humans. The capillary microscopy system consists of a reflected-light microscope with a cold light source, green and infrared filters and a video camera. The video sequences were recorded using the image capture system Framegrabber (Imagenation PXC-200) and a PC (with an Intel Core 2 Duo processor, 1024 MB RAM, 160 GB hard disk, Windows XP Pro), and stored via a DVD recorder (Panasonic LQ-MD800). Quantification of capillary erythrocyte flow velocities was performed using the computer-assisted image analysis system Cap Image Version 8.5 which includes a movie tool as a video sequence storage medium. The method allows estimation of capillary density and tortuosity as well as capillary circulation in the anesthetized pig within a few minutes. First measurements were made after anesthesia induction followed by further measurements during anesthesia maintenance (3 minutes each). No differences in capillary circulation were found. The present method is thus very well suited for long-term microcirculation measurements in pigs, e.g., to evaluate therapeutic interventions in the ischemic limb model.
