Image correlation algorithm for measuring lymphocyte velocity and diameter changes in contracting microlymphatics

Efforts have recently been made to estimate wall shear stress throughout the contractile cycle of mesenteric rat lymphatics with a high speed video microscopy system. This was prompted by reports in the literature that lymphatic pumping is related to wall shear stress. While one can estimate wall shear stress by tracking lymphocyte velocity, it is prohibitively tedious to manually track particles over a reasonable time frame for a good number of experiments. To overcome this, an image correlation method similar to digital particle imaging velocimetry was developed and tested on contracting lymphatics to measure both vessel diameter and fluid velocity. The program tracked temporal fluctuations in spatially averaged velocity with a standard error of prediction of 0.4 mm/s. From these studies we have measured velocities ranging from -2 to 4 mm/s. Diameter changes were also measured with a standard error of 7 microm. These algorithms and techniques could be beneficial for investigating various changes in contractile behavior as a function of changes in velocity and wall shear stress.