LPS-enhanced leukocyte migration assay in 4 dpf zebrafish larvae.

All larvae (nacre) are four days post-fertilization (4 dpf), with anterior to the left, scale bar = 10 μm. A, tail of alive larva without tail cut; B, tail of alive larva with tail cut; C–D, whole-mount MPO staining in uncut tails of zebrafish larvae; C, without lipopolysaccharides (LPS) and D, with lipopolysaccharides (+LPS); E–F, whole-mount MPO staining in cut tails of zebrafish larvae; E, without the inclusion of LPS; F, with the inclusion of LPS. Dark-spots (marked by arrows) represent the migrating leukocytes, which are semi-quantified in the region to the right of the dashed red arc.

Scoring of leukocyte migration in the transected tails of zebrafish larvae.

All larvae (nacre) are four days post-fertilization (4 dpf), with anterior to the left, scale bar = 10 μm. Dark spots (marked by arrows) represent leukocytes migrating to the injured zone in the transected tails. Migrating leukocytes were counted in the region to the right of the dashed red arc. A, tail of an uncut larva; B, tail-cut with score 0; C, tail-cut with score 1; D, tail-cut with score 2; E tail-cut with score 3; F, tail-cut with score 4. ^aExperimental values obtained for each larvae are normalized to a relative value expressed as relative leukocyte migration (RLM); +LPS: with inclusion of lipopolysaccharides. ^bPercentage of anti-inflammatory activity is obtained as (1RLM)x100.