PMID- 8639232
OWN - NLM
STAT- MEDLINE
DCOM- 19960715
LR  - 20191021
IS  - 0939-5075 (Print)
IS  - 0341-0382 (Linking)
VI  - 51
IP  - 3-4
DP  - 1996 Mar-Apr
TI  - Optimization of fast-fluorescence in situ hybridization with repetitive 
      alpha-satellite probes.
PG  - 253-61
AB  - A rapid FISH (fluorescence in situ hybridization) technique (Fast-FISH) for 
      quantitative microscopy has been recently introduced. For highly repetitive DNA 
      probes the hybridization (renaturation) time and the number of necessary washing 
      steps were reduced considerably by omitting formamide or equivalent denaturing 
      chemical agents. Due to low stringency conditions major and minor binding sites 
      of the probes used showed visible FISH signals well suited for quantitative 
      image-microscopy. The discrimination of minor and major binding sites was 
      possible by automated image-processing. Here, a further, quantitative 
      optimization of the Fast-FISH technique is described that allows to clearly 
      discriminate major and minor binding sites of alpha-satellite probes by an easy 
      image classification parameter. With respect to the optimization it was necessary 
      to verify two sensitive parameters (hybridization time and temperature) of the 
      given rapid FISH protocol. As examples the systematic optimization for the two 
      probes D12Z2 (major binding site on the centromere of chromosome 12) and D8Z2 
      (major binding site on the centromere of chromosome 8) are shown. The optimal 
      hybridization conditions concerning rapidness and quality of chromosome 
      morphology were obtained using a hybridization temperature of 70 degrees C and a 
      hybridization time of 60 min. For these conditions major and minor binding sites 
      were clearly discriminated by the intensity maximum Smax of the corresponding 
      FISH-spots.
FAU - Durm, M
AU  - Durm M
AD  - Institut fur Angewandte Physik, Universitat Heidelberg.
FAU - Haar, F M
AU  - Haar FM
FAU - Hausmann, M
AU  - Hausmann M
FAU - Ludwig, H
AU  - Ludwig H
FAU - Cremer, C
AU  - Cremer C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Z Naturforsch C J Biosci
JT  - Zeitschrift fur Naturforschung. C, Journal of biosciences
JID - 8912155
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Satellite)
SB  - IM
MH  - Binding Sites
MH  - Centromere/ultrastructure
MH  - Chromosome Mapping
MH  - Chromosomes, Human, Pair 12
MH  - Chromosomes, Human, Pair 8
MH  - *DNA Probes
MH  - *DNA, Satellite
MH  - Hot Temperature
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Kinetics
MH  - Lymphocytes/cytology
MH  - Metaphase
MH  - Nucleic Acid Denaturation
MH  - *Repetitive Sequences, Nucleic Acid
MH  - Thermodynamics
MH  - Time Factors
EDAT- 1996/03/01 00:00
MHDA- 1996/03/01 00:01
CRDT- 1996/03/01 00:00
PHST- 1996/03/01 00:00 [pubmed]
PHST- 1996/03/01 00:01 [medline]
PHST- 1996/03/01 00:00 [entrez]
AID - 10.1515/znc-1996-3-418 [doi]
PST - ppublish
SO  - Z Naturforsch C J Biosci. 1996 Mar-Apr;51(3-4):253-61. doi: 
      10.1515/znc-1996-3-418.