PMID- 17094016
OWN - NLM
STAT- MEDLINE
DCOM- 20070921
LR  - 20220716
IS  - 1567-2379 (Print)
IS  - 1567-2379 (Linking)
VI  - 38
IP  - 2
DP  - 2007 May
TI  - The specificity of interphase FISH translocation probes in formalin fixed 
      paraffin embedded tissue sections is readily assessed using automated staining 
      and scoring of tissue microarrays constructed from murine xenografts.
PG  - 159-65
AB  - Implementation of interphase fluorescence in situ hybridization (FISH) assays in 
      the clinical laboratory requires validation against established methods. 
      Validation tools in common use include exchange of consecutive sections with 
      another institution that has already established the FISH assay, comparison with 
      conventional banded metaphase cytogenetics, confirmation of specificity using 
      probed normal metaphases, consecutive paraffin sections of a validation set 
      tested by a reference laboratory, and specificity assessment against well 
      characterized cell lines. We have investigated the feasibility of using tissue 
      microarrays (TMA) constructed from murine xenografts as a preliminary 
      specificity-screening tool for validation of interphase FISH assays. Cell lines 
      currently in use for FISH controls are used to generate xenografts in SCID mice 
      which are fixed in formalin and paraffin embedded. A TMA is constructed using 
      duplicate donor cores from the xenograft blocks. Xenografts used represent a wide 
      range of translocations used routinely for formalin fixed paraffin embedded 
      sections evaluated by FISH. Probe cocktails (Abbott-Vysis), for several 
      non-random translocations associated with hematologic neoplasms and soft tissue 
      sarcomas have been used in this manner. On-line deparaffinization, cell 
      conditioning, and prehybridization steps are automated using a staining 
      workstation (Ventana Discovery XT); hybridization and stringency washes are 
      performed manually offline. FISH-probed TMAs are tracked using a Metasystems 
      image scanner and analyzed using classifiers specifically developed for each 
      molecular abnormality. FISH results for each xenograft in the TMA correspond 
      exactly to the genotype previously established for the parent cell line from 
      which the xenograft was prepared. Moderate complexity tissue microarrays 
      constructed from murine xenografts are excellent validation tools for initial 
      assessment of interphase FISH probe specificity.
FAU - Tubbs, Raymond R
AU  - Tubbs RR
AD  - Dept of Clinical Pathology, The Cleveland Clinic Foundation and the Cleveland 
      Clinic Lerner College of Medicine, Cleveland, OH 44195, USA. TubbsR@ccf.org
FAU - Pettay, James
AU  - Pettay J
FAU - Barry, Todd S
AU  - Barry TS
FAU - Swain, Eric
AU  - Swain E
FAU - Loftus, Margaret
AU  - Loftus M
FAU - Cook, James R
AU  - Cook JR
FAU - Skacel, Marek
AU  - Skacel M
FAU - Paine, Gillian
AU  - Paine G
FAU - Roche, Patrick
AU  - Roche P
FAU - Grogan, Thomas
AU  - Grogan T
LA  - eng
PT  - Journal Article
DEP - 20061109
PL  - Netherlands
TA  - J Mol Histol
JT  - Journal of molecular histology
JID - 101193653
RN  - 0 (DNA Probes)
RN  - 1HG84L3525 (Formaldehyde)
SB  - IM
MH  - Animals
MH  - Cell Line, Tumor
MH  - DNA Probes/*genetics
MH  - *Formaldehyde
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - Neoplasms/genetics
MH  - Paraffin Embedding/*methods
MH  - Tissue Array Analysis/*methods
MH  - Xenograft Model Antitumor Assays/*methods
EDAT- 2006/11/10 09:00
MHDA- 2007/09/22 09:00
CRDT- 2006/11/10 09:00
PHST- 2005/10/21 00:00 [received]
PHST- 2006/10/10 00:00 [accepted]
PHST- 2006/11/10 09:00 [pubmed]
PHST- 2007/09/22 09:00 [medline]
PHST- 2006/11/10 09:00 [entrez]
AID - 10.1007/s10735-006-9066-1 [doi]
PST - ppublish
SO  - J Mol Histol. 2007 May;38(2):159-65. doi: 10.1007/s10735-006-9066-1. Epub 2006 
      Nov 9.