FIG. 6.

Quantitative demonstration of lymphangiogenesis following influenza infection. Whole MLN reconstructions of mice administered PBS (left, 0 h) or influenza strain A/H1N1 Puerto Rico 8 (right, 72 hpi). The MLN tissue was thick-sectioned (50 μm), stained with antibodies specific for B220 (red), CD3 (green), and Lyve-1 (blue), imaged by confocal microscopy, and reconstructed by the approach shown in Figure 4. Lymphatic vessels (LVs) were then identified as objects within single xy planes and quantified across a total of (6) nonsequential xy planes for each reconstructed LN. XY planes were analyzed sequentially as outlined in Figure 5 to identify LVs within the T cell cortex. Quantitation of the numbers of identified LVs within each plane is shown with statistical analysis (Student's t test, n=6 XY planes). A color version of this figure is available in the online article at www.liebertpub.com/lrb