What are the steps of immunofluorescence?

1. fixation to preserve morphology and cellular architecture. 2. permeabilization in case plasma membrane is not yet disrupted by fixation, to give antibodies access to intracellular epitopes. 3. embedding in paraffin to fix the tissue (not needed for cells). 4. deparaffination and rehydration. 5. antigen retrieval to restore epitope-antibody reactivity through; - protease induced epitope retrieval; enzymes cleave protein cross-links - heat-induced epitope retrieval; heat and pressure is used 6. blocking to prevent binding of antibodies to non-target epitopes. 7. primary or secondary immunofluorescence. 8. detection using fluorescent microscope.