Figure 2

Data handling workflow. (a) Raw data, demonstrating goals and challenges. Top left: tiled microscopy images show adherent leukocytes , but also have large inhomogeneities due to differences in background illumination between tiles . Additionally, some lanes are blocked and must be excluded from further analysis . Bottom right: the same data after phase congruency calculation. This isolates the important features, while removing effects of background inhomogeneity. (b) Registration of cleaned phase contrast images to the template. Top left: initial manual rigid registration results in mismatches between the phase congruency map (magenta) and the device template (green). Affine registration using Mattes mutual information results in near-perfect alignment between the data and the template (bottom right). (c) Raw data from cells run in the presence of RBCs (green) superimposed with the device template (magenta), demonstrating the quality of the resultant alignment. Wall-adherent and rolling leukocytes colocalize with the wall delineated on the template. (d) Example of ROI from media (LCIS) condition. (e) Example of ROI from RBC condition, demonstrating sidewall-adherent leukocytes, a center adherent leukocyte, flowing leukocytes, and rolling leukocytes.