6A Method for Analysis of Cell Morphology, Banded Karyotype, and Immunoperoxidase Identification of Lymphocyte: Subset on the Same Cell

Abstract

The authors describe a method that allows three staining pro-cedures to be applied to the same mitotic cell, namely Giemsa staining for morphology, immunoperoxidase staining for surface markers, and banding of the chromosomes for karyotype. After culturing of the blood lymphocytes, colchicine was added to arrest the cells in mitosis. After mild hypotonic treatment, the cells were cytocentrifuged to glass slides. The mitotic cells of different lymphatic cell subsets were identified with monoclonal anti-sera against T-helper-, T-suppressor-, and B-cells using the immu-noperoxidase technic. The cells were counterstained with Giemsa for morphologic identification. After removal of Giemsa stain and peroxidase, the chromosomes were G-banded.