Identification of Resting Human T and B Lymphocytes by Acid α‐Naphthyl Acetate Esterase Staining Combined with Rosette Formation with Staphylococcus aureus Strain Cowan 1

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Abstract
We describe a method enabling the identification of both lymphocyte class and morphology from a single microscopic slide. As a marker for B cells we used surface immunoglobulin. The surface‐Ig‐carrying cells were rosetted after polyvalent anti‐Ig treatment with Staphylococcus aureus strian Cowan 1 (StaCw) and the cells were cytocentrifuged onto a microscope slide. The lymphocytes forming rosettes with StaCw were identical with cells expressing surface Ig studied by fluorescein‐isothiocyanate‐conjugated anti‐Ig. As a marker for T cells we used the acid α‐naphthyl acetate esterase (ANAE) histochemical marker. The cell smears were first stained for ANAE and subsequently counterstained to distinguish also cell morphology. The ANAE‐marker‐carrying cells were all included in the population of lymphocytes forming rosettes with sheep erythrocytes. Thus both T and B lymphocytes could be simultaneously identified from a single microscopic slide, and we therefore recommend the method for routine clinical work.