Micro-PTV measurement of the fluid shear stress acting on adherent leukocytes in vivo.

Leukocyte adhesion is determined by the balance between molecular adhesive forces and convective dispersive forces. A key parameter influencing leukocyte adhesion is the shear stress acting on the leukocyte. This measure is indispensable for determining the molecular bond forces and estimating cell deformation. To experimentally determine this shear stress, we used microparticle tracking velocimetry analyzing more than 24,000 images of 0.5 microm fluorescent microbeads flowing within mildly inflamed postcapillary venules of the cremaster muscle in vivo. Green fluorescent protein, expressed under the lysozyme-M promoter, made leukocytes visible. After applying stringent quality criteria, 3 of 69 recordings were fully analyzed. We show that endothelial cells, but not leukocytes, are covered by a significant surface layer. The wall shear rate is nearly zero near the adherent arc of each leukocyte and reaches a maximum at the apex. This peak shear rate is 2-6-fold higher than the wall shear rate in the absence of a leukocyte. Microbead trajectories show a systematic deviation toward and away from the microvessel axis upstream and downstream from the leukocyte, respectively. The flow field around adherent leukocytes in vivo allows more accurate estimates of bond forces in rolling and adherent leukocytes and improved modeling studies.