Describe both direct and indirect antigen staining in flow cytometry and the advantages
and disadvantages of each.
What will be an ideal response
In direct staining, a monoclonal antibody is labeled with a fluorochrome and then reacted with a cell. In indirect
staining, an unlabeled monoclonal antibody is reacted with a cell. Then, a second labeled antibody directed against
the species that produced the first antibody is added. For example, anti-CD4 of mouse origin is reacted with a cell
and then fluorescein-labeled anti-mouse globulin is added. Direct staining is faster, but antibodies must be
available that have labels. If unlabeled antibodies are the only reagent available, then the indirect technique renders
them useful. In addition, indirect staining increases the ability to detect sparsely expressed markers. The
disadvantages, though, are increased nonspecific staining and a more lengthy procedure.
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