Images of hemocytes and average proportional distribution of circulating hemocytes in naïve and challenged larvae.

<p>Confocal images of hemocytes stained with DAPI (blue) for nuclei (N) and with filamentous actin (F-actin; red) for cytoskeleton visualization. Hemocytes were classified as granulocytes (A and G), plasmatocytes (B and H), oenocytoids (C and I), spherulocytes (D and J), prohemocytes (E and K), and adipohemocytes (F and L) on the basis of their size, morphology, and dye-staining properties. Pseudopodia or filopodia on the plasma membrane, are indicated by white arrow (A, B, G, and H). The panels M and N show melanized oenocytoids (M) and granulocyte (N). Hemocyte counting was performed by differential hemocyte count (DHC) and 35 larvae (11,235 hemocytes; 321 hemocytes per larva) were used to determine the percentage of the six circulating hemocyte types; five naïve larvae and three challenged larvae at each time point (4, 8, 12, 24, and 48 h h) post infection. Results are given as means and standard deviation. *(P<0.05). Challenged larvae were infected with <i>E</i>. <i>coli</i> (panel O) or <i>S</i>. <i>cerevisiae</i> (panel P) whereas naïve larvae were injected with sterile saline solution. PR, prohemocytes; PL, plasmatocytes; GR, granulocytes; SP, spherulocytes; OE, oenocytoids; AD, adipohemocytes. Scale bar = 20 µm.