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Williams Hematology > Part XI. Transfusion Medicine > Color Atlas XXIII: Fluorescence In Situ Hybridization (FISH) for Cytogenetic Analysis >

Fluorescence In Situ Hybridization (FISH) for Cytogenetic Analysis

Plate XXIII

Fluorescence in situ hybridization and spectral karyotyping analysis of hematologic malignant diseases. Panels B, D, and E illustrate images of metaphase and interphase cells following FISH; the cells are counterstained with 4,6-diamidino-2-phenylindole-dihydrochloride (DAPI). (A) Schematic diagram of the BCR and ABL loci, location of the BCR and ABL dual fusion probe (Vysis, Inc), and configuration of signals in interphase cells. (B) Hybridization of the BCR-ABL dual fusion probe to metaphase and interphase cells with the t(9;22). In cells with the t(9;22), only one green and one red signal is observed on the normal 9 and 22 homologs, and two yellow fusion signals (arrows) are observed on the der(9) and the der(22) (Ph chromosome) chromosomes as a result of the juxtaposition of the ABL and BCR sequences. (C) Schematic diagram of the MLL gene, location of the MLL break-apart probe (Vysis Inc.), and configuration of signals in interphase cells. (D) Hybridization of the MLL break-apart probe to metaphase and interphase cells with a t(11q23). In cells with MLL translocation, a yellow fusion signal is observed for the germline configuration on the normal chromosome 11 homologue, a green signal is observed in the der(11) chromosome, and a red signal is observed on the partner chromosome. (E) Hybridization of directly-labeled centromere-specific probes for the X (CEPX Spectrum Orange, Vysis Inc.) and Y (CEPX Spectrum Green, Vysis Inc.) chromosomes to interphase cells from a marrow aspirate of a female patient with AML who received a marrow transplant from a male donor. Centromere-specific probes hybridize to the repetitive DNA sequences that are present at the centromeres of human chromosomes. (F) Spectral karyotyping analysis of a metaphase cell from an AML-M7. Twenty-four differentially labeled probes representing each human chromosome are co-hybridized, and imaging analysis software assigns a unique color to each. A complex karyotype was identified by conventional cytogenetic analysis, including a derivative chromosome 1 with additional material of unknown origin on 1p, a deletion of 8p, a derivative chromosome 11 resulting from an unbalanced translocation involving 1 and 11, and a derivative chromosome 12, consisting of 11q and 12q. The results of spectral karyotyping confirmed the identity of the rearranged chromosome 12 (arrowhead), but clarified the other abnormalities. The additional material on 1p was derived from chromosome 8 (long arrow, blue signal), and the der(11) actually consisted of material from chromsomes 1, 11, and 12 (short arrow, 11p white signal; chromosome 12 brown signal; 1p blue-pink signal).

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