-immunize mouse with antigen A

-euthanize mouse

-harvest spleen B cells which are producing antibody

-fuse spleen cells with myeloma cells to create a hybridoma

-transfer to cell culture medium

-select for fused cells using HGPRT

-screen hybridoma antibodies

-grow antigen A specific hybridomas for monoclonal antibody production

-label with fluorescent molecules

-polyelthylene glycol used to create fusion

-spleen cells produce antibody

-myeloma cell creates immortality

-Fluorescence-activated cell sorting

-cells sorted by particle size

-cell surface markers detected via fluorophore conjugation

-internal cell attributes are detected via fluorescent indicator

-size

-complexity

-phenotype

-health

-forward scatter: particle size

-side scatter: granularity

-fluorescence