Two new applications of the immunoperoxidase method: cell-by-cell quantitation of surface immunoglobulins and automated recognition of B-lymphocytes.

Two new applications of the immunoperoxidase method are reported. In the first one, cells after incubation with antibodies coupled with peroxidase washing and staining by carbazol are cytocentrifuged and individually analyzed for their content on surface immunoglobulin (Ig) in two photometric microscopes. The results included the following: 1. Maturation of mice B-lymphocytes during ontogeny is accompanied by the appearance of a population containing intermediate to high amounts of SIg. 2. The CLL B-lymphocyte pattern as compared with that of normal B-lymphocytes shows a decrease of SIg and a homogeneous distribution. In the second application, mice splenic lymphocytes are incubated with antimouse Ig coupled with peroxidase and analyzed through Hemalog D. B-lymphocytes are separated from T-lymphocytes in a distinct population since they become positive for peroxidase.