PMID- 18274524
OWN - NLM
STAT- MEDLINE
DCOM- 20081110
LR  - 20161124
IS  - 1750-2799 (Electronic)
IS  - 1750-2799 (Linking)
VI  - 3
IP  - 2
DP  - 2008
TI  - Fluorescence and chromogenic in situ hybridization to detect genetic aberrations 
      in formalin-fixed paraffin embedded material, including tissue microarrays.
PG  - 220-34
LID - 10.1038/nprot.2007.534 [doi]
AB  - Screening for specific genetic aberrations by fluorescence and chromogenic in 
      situ hybridization (fluorescence in situ hybridization (FISH) and chromogenic in 
      situ hybridization (CISH)) can reveal associations with tumor types or subtypes, 
      cellular morphology and clinical behavior. FISH and CISH methodologies are based 
      on the specific annealing (hybridization) of labeled genomic sequences (probes) 
      to complementary nucleic acids within fixed cells to allow their detection, 
      quantification and spatial localization. Formalin-fixed paraffin embedded (FFPE) 
      material is the most widely available source of tumor samples. Increasingly, 
      tissue microarrays (TMAs) consisting of multiple cores of FFPE material are being 
      used to enable simultaneous analyses of many archival samples. Here we describe 
      robust protocols for the FISH and CISH analyses of genetic aberrations in FFPE 
      tissue, including TMAs. Protocols include probe preparation, hybridization and 
      detection. Steps are described to reduce background fluorescence and strip probes 
      for repeat FISH analyses to maximize the use of tissue resources. The basic 
      protocol takes 2-3 d to complete.
FAU - Summersgill, Brenda
AU  - Summersgill B
AD  - Molecular Cytogenetics, The Institute of Cancer Research, 15 Cotswold Road, 
      Sutton, Surrey SM2 5NG, UK.
FAU - Clark, Jeremy
AU  - Clark J
FAU - Shipley, Janet
AU  - Shipley J
LA  - eng
PT  - Journal Article
PL  - England
TA  - Nat Protoc
JT  - Nature protocols
JID - 101284307
RN  - 0 (Chromogenic Compounds)
RN  - 0 (ERG protein, human)
RN  - 0 (Proto-Oncogene Proteins c-myc)
RN  - 0 (Trans-Activators)
RN  - 0 (Transcriptional Regulator ERG)
RN  - 1HG84L3525 (Formaldehyde)
RN  - EC 3.4.21.- (Serine Endopeptidases)
RN  - EC 3.4.21.- (TMPRSS2 protein, human)
SB  - IM
MH  - Chromogenic Compounds
MH  - *Chromosome Aberrations
MH  - Formaldehyde
MH  - In Situ Hybridization/*methods
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Paraffin Embedding
MH  - Proto-Oncogene Proteins c-myc/analysis
MH  - Rhabdomyosarcoma/genetics
MH  - Serine Endopeptidases/analysis
MH  - Tissue Array Analysis/methods
MH  - Tissue Fixation
MH  - Trans-Activators/analysis
MH  - Transcriptional Regulator ERG
EDAT- 2008/02/16 09:00
MHDA- 2008/11/11 09:00
CRDT- 2008/02/16 09:00
PHST- 2008/02/16 09:00 [pubmed]
PHST- 2008/11/11 09:00 [medline]
PHST- 2008/02/16 09:00 [entrez]
AID - nprot.2007.534 [pii]
AID - 10.1038/nprot.2007.534 [doi]
PST - ppublish
SO  - Nat Protoc. 2008;3(2):220-34. doi: 10.1038/nprot.2007.534.