Clone rb6 8c5

Microspheres consisting of 1:1 collagen and alginate of approximately 30-200 µm in diameter were centrifuged, the supernatant was removed and 100 µl of the spheres themselves were injected subcutaneously into 12, approximately 24-week-old, female C57Bl/6 mice, in a total of three technical replicates, using G27 needles on a 1 ml syringe. Five days after injection the mice were killed and the hydrogel microsphere plaques were excised and fixed in 4% PFA at 4°C overnight. Plaques were stained according to a previously described protocol with an anti-mouseF4/80 antibody (1:200, Clone CI:A3-1, AbD Serotec, Bio-Rad, Cat#: MCA497GA) to visualize macrophages, or an anti-mouseLy6G antibody (1:200, Clone RB6-8C5, Abcam, Cat#: ab25377) to visualize neutrophils, coupled to secondary goat-anti-rat-Cy5 (Millipore) or goat-anti-rabbit-Cy3 (Millipore) antibodies. The antibodies have been validated in the scientific literature as well as by the manufacturers. Animal studies were approved by the North Stockholm Research Animal Ethical Council.