Interactions of leukocytes with vessel walls and with other blood cells, studied by high-resolution intravital videomicroscopy of spleen.

Visualization of circulating leukocytes in vivo is difficult because their optical density differs so little from that of plasma. We have obtained intravital microscopic images of leukocytes at high resolution in spleens of rats and mice by means of an inverted microscope with a 100 x oil-immersion lens; oblique lighting gave improved contrast. Photographic evidence and quantitative data describing the behaviour of lymphocytes, polymorphonuclear leukocytes (PMNs), and macrophages within the microvasculature are presented. Mean numbers of marginated lymphocytes in venous vessels ranged from 0.1 to 4.5 per 1000 microns 2 of wall surface, and speeds of rolling from 11 to 20 microns/sec. Adherence times of leukocytes to vessel walls were log normally distributed, median values being 30, 130, and 560 sec, for lymphocytes, PMNs, and macrophages, respectively. Mean speeds of migration along luminal surfaces were similar (7-19 microns/min) for all three types of cells. Lymphocyte migration outward through the venular wall, observed on two occasions, took 1-2 min. Median values for duration of adherence of RBCs and lymphocytes to macrophages were 1 and 42 sec, respectively. Phagocytosis of a lymphocyte was observed and took 3 min. Macrophages often underwent dramatic changes in shape, including formation of a pseudopod up to 155 microns in length. High-resolution intravital videomicroscopy of spleen has great potential for studying leukocyte behaviour, e.g., homing and migration of lymphocytes, and immunologically related macrophage-lymphocyte interactions in vivo.