PMID- 7981136
OWN - NLM
STAT- MEDLINE
DCOM- 19950105
LR  - 20061115
IS  - 0921-8912 (Print)
IS  - 0921-8912 (Linking)
VI  - 7
IP  - 1
DP  - 1994 Jul
TI  - Improvements in visualisation and localisation of human papillomavirus DNA in 
      CaSki cells by fluorescence in situ hybridization, laser scanning confocal 
      microscopy and three-dimensional image reconstruction.
PG  - 53-61
AB  - The visual interpretation and localisation of specific DNA sequences in three 
      dimensions in cell nuclei was investigated by fluorescence in situ hybridization 
      (FISH) and laser scanning confocal microscopy (LSCM) using CaSki cells containing 
      600 copies per cell of human papillomavirus (HPV) DNA type 16 integrated in 
      cellular DNA. Biotinylated DNA probes were used and DNA-DNA hybrids were revealed 
      by a three-step reaction involving a rabbit anti-biotin antibody, a biotinylated 
      goat anti-rabbit antibody and a streptavidin-fluorescein isothiocyanate complex. 
      The DNA from cell nuclei was counterstained with propidium iodide. With standard 
      fluorescence microscopy, some dense fluorescent spots were seen in the cell 
      nuclei. Similarly, with LSCM, some hybridization spots were observed in the cell 
      nuclei but they were at different levels of the nuclei as shown by successive 
      nuclear sections taken along the z axis. The visualisation of multiple 
      hybridization spots confirmed the presence of multiple integration sites of HPV 
      16 DNA in CaSki cells. Association of LSCM with three-dimensional reconstructions 
      lead to spatial images of hybridization spots obtained by stacking (x,y) images 
      from consecutive confocal planes. Rotation of the reconstructed cell nuclei 
      around the y axis makes it possible to distinguish closely adjacent spots. The 
      combination of these techniques improves the detection of hybridization spots and 
      may be of interest to further determine whether the HPV DNA is episomal or 
      integrated in infected cells.
FAU - Lizard, G
AU  - Lizard G
AD  - Centre Commun de Cytometrie en Flux-INSERM U80, Hopital E. Herriot, Lyon, France.
FAU - Usson, Y
AU  - Usson Y
FAU - Chignol, M C
AU  - Chignol MC
FAU - Chardonnet, Y
AU  - Chardonnet Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Anal Cell Pathol
JT  - Analytical cellular pathology : the journal of the European Society for 
      Analytical Cellular Pathology
JID - 8911016
RN  - 0 (DNA, Viral)
SB  - IM
EIN - Anal Cell Pathol 1995 Jun;8(4):357-8
MH  - DNA, Viral/*analysis
MH  - Humans
MH  - Image Processing, Computer-Assisted
MH  - In Situ Hybridization, Fluorescence
MH  - Microscopy, Confocal
MH  - Papillomaviridae/*isolation & purification
MH  - Tumor Cells, Cultured/*virology
EDAT- 1994/07/01 00:00
MHDA- 1994/07/01 00:01
CRDT- 1994/07/01 00:00
PHST- 1994/07/01 00:00 [pubmed]
PHST- 1994/07/01 00:01 [medline]
PHST- 1994/07/01 00:00 [entrez]
PST - ppublish
SO  - Anal Cell Pathol. 1994 Jul;7(1):53-61.