PMID- 9313806
OWN - NLM
STAT- MEDLINE
DCOM- 19971023
LR  - 20170214
IS  - 0022-1554 (Print)
IS  - 0022-1554 (Linking)
VI  - 45
IP  - 10
DP  - 1997 Oct
TI  - Sensitive multicolor fluorescence in situ hybridization using catalyzed reporter 
      deposition (CARD) amplification.
PG  - 1439-46
AB  - We describe the simultaneous localization of DNA sequences in cell and chromosome 
      preparations by means of differently fluorochrome-labeled (AMCA, FITC, TRITC) 
      tyramides using the catalyzed reporter deposition (CARD) procedure. For this 
      purpose, repeated as well as single-copy DNA probes were labeled with biotin, 
      digoxigenin, and FITC, hybridized, and visualized with three different 
      cytochemical detection systems based on horseradish peroxidase conjugates. These 
      were sequentially applied to interphase nuclei and metaphase chromosomes at low 
      concentrations to prevent crossreaction and nonspecific background. In situ 
      localized peroxidase activity was visualized by the deposition of 
      fluorochrome-labeled tyramide molecules. To allow specific deposition of a second 
      and a third tyramide conjugate for multiple-target fluorescence in situ 
      hybridization (FISH), remaining peroxidase activity was always completely 
      inactivated by a mild acid treatment before application of the next peroxidase 
      conjugate. The CARD reactions were optimized for maximal signal-to-noise ratio 
      and discrete localization by tuning reaction time, H2O2, and tyramide 
      concentrations. For both repeated and single-copy DNA targets, high FISH signal 
      intensities were obtained, providing improvement of sensitivity over conventional 
      indirect detection systems. In addition, the fluorescence CARD detection system 
      proved to be highly efficient and easy to implement in multiple-labeling studies, 
      such as reported here for FISH.
FAU - Speel, E J
AU  - Speel EJ
AD  - Department of Molecular Cell Biology & Genetics, University of Maastricht, The 
      Netherlands.
FAU - Ramaekers, F C
AU  - Ramaekers FC
FAU - Hopman, A H
AU  - Hopman AH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Histochem Cytochem
JT  - The journal of histochemistry and cytochemistry : official journal of the 
      Histochemistry Society
JID - 9815334
RN  - 0 (DNA Probes)
RN  - 0 (Fluorescent Dyes)
RN  - 9007-49-2 (DNA)
RN  - EC 1.11.1.- (Horseradish Peroxidase)
RN  - X8ZC7V0OX3 (Tyramine)
SB  - IM
MH  - Carcinoma, Transitional Cell/chemistry/genetics
MH  - Centromere/*chemistry
MH  - DNA/*analysis
MH  - DNA Probes
MH  - Fluorescent Dyes/*chemistry
MH  - Gene Expression
MH  - Horseradish Peroxidase
MH  - Humans
MH  - *Immunoenzyme Techniques
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Metaphase
MH  - Tumor Cells, Cultured
MH  - Tyramine/*chemistry
EDAT- 1997/10/06 00:00
MHDA- 1997/10/06 00:01
CRDT- 1997/10/06 00:00
PHST- 1997/10/06 00:00 [pubmed]
PHST- 1997/10/06 00:01 [medline]
PHST- 1997/10/06 00:00 [entrez]
AID - 10.1177/002215549704501013 [doi]
PST - ppublish
SO  - J Histochem Cytochem. 1997 Oct;45(10):1439-46. doi: 10.1177/002215549704501013.