Morphology antibody chromosome technique for determining phenotype and genotype of the same cell.
The morphology antibody chromosome (MAC) technique is a powerful combination of methods that permits analysis of both phenotype and genotype on a single interphase or mitotic cell as a basis for lineage analysis of neoplastic and normal cells. This unit describes MAC with sequential phenotypic analysis using antibody and an alkaline phosphatase anti-alkaline phosphatase (APAAP) complex and genotypic analysis using in situ hybridization with either enzymatic or fluorescence detection. Alternate methods for phenotypic analysis are also described, which include the use of horseradish peroxidase-conjugated antibodies, fluorochrome-conjugated antibodies, May-Grunwald-Giemsa cytochemical staining, and Sudan black B cytochemical staining. An additional protocol describes G- and C-banding as alternatives to in situ hybridization (ISH) for genotyping MAC specimens. Support protocols describe methods for preparing specimenscytospin preparations, in situ cultures, paraffin-embedded or cryostat sections, and blood and bone marrow smears. Basic Protocol 2 describes a procedure for chromosome painting of previously GTG-banded slides.
Knuutila
Sakari
S
University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland.
D002871:Chromosome Banding; D002877:Chromosomes, Human; D005838:Genotype; D006801:Humans; D017403:In Situ Hybridization; D010641:Phenotype