I would like to know an experiment to detect phagocytosis by raw 264.7 cells on infection with influenza virus. I used Vybrant phagocytosis assy kit and do not see any phagocytosis both in cells infected with IAV and cells infected with IAV and stimuated using LPS.
To see a nice phagocytosis with macrophages they can be be primed during 1 to 3 h with LPS in adherence. Then they can be stimulated with opsonized particles (IgG or AB serum) for 1 h. Opsonization is an important step to improve specific phagocytosis.The phagocytosis assay can be made also with macrophages in suspension and then the phagocytosis is measured by FACS after trypan blue treatment or activated cells can be deposited on a slide to adhere and observed under microscope.
Thank you for the reply. I am looking at the cells during infection. So, basically studying the infected cells and the course of phagocytosis. Is there any experiment design to study?
Indeed basically you can follow the phagocytosis of the infected cells by FACS or directly by observing them under fluorescence microscope at different times of contact with bacteria. You can test another bacteria more pathogenic as St aureus or opsonized zymosan (yeast) or opsonized fungi like C. albicans.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus