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1. Cell Biology Lab 6Phagocytosis PCB 4023 L
TA: Sushmita Mustafi
2. Objectives: Phagocytosis.
Rab proteins
Role of Rab5 in phagocytosis
Rab5 Effector.
Experimental design.
3. Endocytosis
5. Rab proteins and vesicle transport
6. Rab GTPase protein
7. Membrane fusion
8. Phagocytic index a measure of phagocytic activity
Determined by counting the number of bacteria ingested per phagocyte during a limited period of incubation of a suspension of bacteria and phagocytes.
9. Seeding cells….
10. Experimental design 1. Incubate cells with phagocytic particles
Cells : Eclone/ RAB5 eclone.
Incubation medium: HBSS-BSA
2. Initiate phagocytosis : By putting the cells in 37c incubator.
3. Stop phagocytosis: Adding fixative to the cells. Paraformaldehyde.
4. Make the fixed cells permiable to stain:
Add tritonX to cells
5. Stain the nucleus of the cells: stain with DAPI.
11. Mounting slides