Fluorescence In Situ Hybridization

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Amy Y-Y Chen, MD Andrew Chen, MD

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Fluorescence In Situ Hybridization Amy Y.-Y. Chen, Andrew Chen Journal of Investigative Dermatology Volume 133, Issue 5, Pages 1-4 (May 2013) DOI: 10.1038/jid.2013.120 Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Schematic diagram of the fluorescence in situ hybridization (FISH) technique. Reprinted from O’Connor, 2008 Journal of Investigative Dermatology 2013 133, 1-4DOI: (10.1038/jid.2013.120) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Histology and dual-target fluorescence in situ hybridization (FISH) with probe RMC11B022 for chromosome 11p and RMC11P008 for chromosome 11q. (a) Case 2, normal CGH measurement; (b) case 13, gain of chromosome 11p; and (c) case 15, normal CGH measurement. FISH shows that infrequent cells with large nuclei (arrow) are polyploid; green signals, chromosome 11p; red signals, chromosome 11q. Reprinted with permission from Bastian et al., 1999 Journal of Investigative Dermatology 2013 133, 1-4DOI: (10.1038/jid.2013.120) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 IRF4 locus-specific fluorescence in situ hybridization (FISH) strategy. (a) Schematic representation of all BAC clones hybridizing to the 6p25 region used in this study. RP11-119L15, CTD-2317K17, and CTD-3052J12 BAC clones, which showed nonspecific hybridization on chromosome 16, were discarded. The break-apart BAC probe strategy used CTD-2308G5 as the 5 IRF4 BAC probe and RP11-164H16 as the 3 IRF4 BAC probe. (b) The FISH signal pattern expected in interphase nuclei samples. Normal nuclei would exhibit a two-fusion (2F) signal pattern corresponding to the juxtaposition of BAC clones probes. Nuclei with an IRF4 locus break point, suggesting translocation, would show a split signal pattern (1F–1R–1G). Nuclei with extra copies of a nonrearranged IRF4 locus should exhibit more than a 2F signal pattern. Reprinted with permission from Pham-Ledard et al., 2010. Journal of Investigative Dermatology 2013 133, 1-4DOI: (10.1038/jid.2013.120) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Histological, immunophenotypical, and fluorescence in situ hybridization (FISH) aspects of three typical cases with and without IRF4 locus rearrangement. Left: Lymph node section of cutaneous anaplastic large-cell lymphoma (C-ALCL) without IRF4 locus rearrangement. (a) Hematoxylin–eosin and safran (HES). Bar = 20 μm. (b) Positivity of CD30 immunostaining. (c) Immunostaining shows expression of multiple myeloma antigen 1 (MUM1) by more than 85% of tumor cells. (d) Normal FISH signal pattern (2F). Bar = 5 μm. Middle: Skin section of case 6 with C-ALCL and IRF4 rearrangement. (e) HES, × 400. (f) Positivity of CD30 immunostaining. (g) Immunostaining shows MUM1 expression by more than 85% of tumor cells. (h) Split FISH signal pattern (1F–1R–1G). Bar = 5 μm. Right: Skin section of case 8 with transformed mycosis fungoides and IRF4 locus rearrangement. (i) HES, × 400. (j) Positivity of CD30 immunostaining. (k) Immunostaining shows MUM1 expression by 10–50% of large tumor cells. (l) FISH signal pattern shows an extra signal of SpectrumGreen-labeled RP11-164H16 (2F+1G extra signal), indicating that the break point maps to the 6p25 region in the genomic area corresponding to the RP11-164H16 sequence. Bar = 5 μm. Reprinted with permission from Pham-Ledard et al., 2010. Journal of Investigative Dermatology 2013 133, 1-4DOI: (10.1038/jid.2013.120) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Journal of Investigative Dermatology 2013 133, 1-4DOI: (10. 1038/jid Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Journal of Investigative Dermatology 2013 133, 1-4DOI: (10. 1038/jid Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions