ExerciseⅤ Phagocytosis of Macrophage
Macrophage is primary cell of the mononuclear phagocyte system.The main function of Macrophage is to dispose of microbes and dead cells through the process of phagocytosis.It is important in the innate immune system and adaptive immune system.It can be stimulated by antigen，and then activate.Here，we introduce a method to measure the ability of macrophage.
Phagocytosis of Macrophage—in vivo
Materials
1.Mouse.
2.Bacterial suspension of Staphylococcus（1×107 cells/ml）.
3.6%sterile starch solution.
4.Wright's reagent.
Methods
1.Inject 1ml of 6%sterile starch solution intraperitoneally to mouse.
2.72 h later，inject 1ml of bacterial suspension of Staphylococcus，massage softly the abdomen to ensure spread of bacterial.
3.After 30～40 min，draw neck to kill mouse，cut open the skin and peritoneum to expose the peritoneal cavity.
4.Draw a little of fluid and place on a slide，smear symmetrically and stain with Wright's reagent.
5.Wright's reagent stain add a little drop of wright's reagent，stain for 1 min and then add the same amount of 0.9% NaCl solution on it.Shake it，gently，stain 5 min.(https://www.daowen.com)
6.Dry with bibulous paper，observe under microscope to see.
Phagocytosis of Macrophage—in vitro
1.Inject 1ml of 6%sterile starch solution intraperitoneally to mouse.
2.After 72 h，draw neck to kill mouse，cut open the skin and peritoneum to expose the peritoneal cavity.
3.Draw the fluid from peritoneal cavity，centrifuge 5 min at 1000r/min，and discard supernatant.
4.Wash macrophage with Hank's 5 min at 1000r/min.
Repeat once.
5.Count cells and adjust concentration to 1×106 cells/ml with Hank's.
6.Add equal volume macrophage suspension and bacterial suspension of Staphylococcus to tube.
7.Incubate at 37℃for 30 min（shake two times），centrifuge 5 min at 500r/min，and discard supernatant.
8.Shake tube slightly，place a drop of supernatant on a slide and mix with a drop of wright's.
9.Observe under microscope.