Kinetics of red blood cell passage through interendothelial slits into venous sinuses in rat spleen, analyzed by in vivo microscopy

Abstract

Sequential photomicrographs of RBCs passing through interendothelial slits (IES) in walls of venous sinuses in rat spleen were obtained by video recording in vivo microscopic views. Kinetics of RBC passage were analyzed by slow-motion playback of recordings. An inverted microscope and oblique lighting from a water-cooled fiber optic light source were key elements in obtaining images of sufficient quality for analysis. The direction of RBC passage through IES was, invariably, from reticular spaces of the red pulp into venous sinuses. RBC flow through an individual IES occurred as brief discontinuous bursts, separated by periods of zero, or near zero, flow. Mean rates of RBC flow through six IES analyzed in normal relaxed spleen ranged from 1.4 to 9.1 cells/15 sec, the total RBCs studied being 1523 and the total combined period of observation 98 min. The maximum instantaneous rate was 10 RBCs/sec. RBC transit times ranged from 0.02 to 60.5 sec, even for a single IES; the distribution was highly skewed: median 0.23 sec, mean 1.7 sec. Analysis of RBC flow through two closely adjacent IES simultaneously, for 30 min, showed that most bursts were asynchronous. The results indicate that changes in caliber of IES are primarily responsible for the observed pattern of flow. It was estimated that only 19% of the total IES present anatomically actually allowed passage of RBCs during any 5-min period.