Unit

UNIT 4.7 Morphology Antibody Chromosome Technique for Determining Phenotype and Genetic Status of the Same Cell

  1. Sakari Knuutila1,
  2. Satu Mustjoki2

Published Online: 1 JUL 2012

DOI: 10.1002/0471142905.hg0407s74

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Knuutila, S. and Mustjoki, S. 2012. Morphology Antibody Chromosome Technique for Determining Phenotype and Genetic Status of the Same Cell. Current Protocols in Human Genetics. 74:4.7:4.7.1–4.7.43.

Author Information

  1. 1

    University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland

  2. 2

    Department of Medicine, Division of Hematology, Helsinki University Central Hospital, Helsinki, Finland

Publication History

  1. Published Online: 1 JUL 2012

Abstract

The morphology antibody chromosome (MAC) technique is a combination of methods that permits analysis of both phenotypic and genetic evaluation 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 specimens, cytospin preparations, in situ cultures, paraffin-embedded or cryostat sections, and blood and bone marrow smears. Also described is a procedure for chromosome painting of previously GTG-banded slides. An additional protocol is included for FISH analysis on sorted hematopoietic stem cells and its application in the detection of leukemic stem cells. For overcoming the drawbacks of scarcity and variability from case to case of malignant plasma cells in multiple myeloma FISH analyses, a protocol is included for the enrichment of plasma cells by immunomagnetic beads. Curr. Protoc. Hum. Genet. 74:4.7.1-4.7.43. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • lineage specificity;
  • immunophenotype;
  • genomic alterations;
  • chromosomes;
  • fluorescence in situ hybridization;
  • hematopoietic stem cells;
  • multiple myeloma