Dorsal skinfold chamber technique for intravital microscopy in nude mice

Abstract

For many years, observation chambers, implanted in various animal species and in man, have been used for intravital microscopy of the microcirculation in granulation tissue, preformed tissue, or of the microvascularization of tissue implants. We describe herein a skinfold chamber model for the intravital microscopic investigation of striated skin muscle in immunoincompetent, nude mice over a minimum period of 2 weeks. Using fluorescent markers for contrast enhancement of plasma (fluorescein isothiocyanate-dextran) and leukocytes (acridine orange), the presented model allows the quantitative analysis of 1) the microhemodynamic parameters microvessel diameter and red blood cell velocity in arterioles (16 to 50 mu diameter), capillaries (4 to 9 mu diameter), and post-capillary venules (19 to 60 mu diameter), 2) leukocyte/endothelium interaction in these vessel segments, 3) functional capillary density and intercapillary distance, and 4) endothelial cell integrity. These parameters can be assessed in the microcirculation of the striated muscle tissue under normal or pathological conditions, as well as in the microcirculation of transplanted xenogeneic (human) neoplastic and non-neoplastic tissue grafts.