PMID- 7928414
OWN - NLM
STAT- MEDLINE
DCOM- 19941104
LR  - 20190830
IS  - 0301-5564 (Print)
IS  - 0301-5564 (Linking)
VI  - 101
IP  - 4
DP  - 1994 Apr
TI  - Laser scanning confocal microscopy and quantitative microscopy with a charge 
      coupled device camera improve detection of human papillomavirus DNA revealed by 
      fluorescence in situ hybridization.
PG  - 303-10
AB  - Epithelial cervical CaSki, SiHa and HeLa cells containing respectively 600 copies 
      of human papillomavirus (HPV) DNA type 16, 1-2 copies of HPV DNA type 16 and 
      10-50 copies of HPV DNA type 18 were used as model to detect different quantities 
      of integrated HPV genome. The HPV DNA was identified on cell deposits with 
      specific biotinylated DNA probes either by enzymatic in situ hybridization (EISH) 
      or fluorescence in situ hybridization (FISH) involving successively a rabbit 
      anti-biotin antibody, a biotinylated goat anti-rabbit antibody and 
      streptavidin-alkaline phosphatase complex or streptavidin-fluorescein 
      isothiocyanate complex. With brightfield microscopy and EISH, hybridization spots 
      were observed in CaSki and HeLa cells but hardly any in SiHa cells. With 
      fluorescence microscopy and FISH, hybridization spots were clearly seen only on 
      CaSki cell nuclei. In an attempt to improve the detection of low quantities of 
      HPV DNA signals revealed by FISH, laser scanning confocal microscopy (LSCM) and 
      quantitative microscopy with an intensified charge coupled device (CCD) camera 
      were used. With both LSCM and quantitative microscopy, as few as 1-2 copies of 
      HPV DNA were detected and found to be confined to cell nuclei counterstained with 
      propidium iodide. Under Nomarski phase contrast, a good preservation of the cell 
      structure was observed. With quantitative microscopy, differences in the number, 
      size, total area and integrated fluorescence intensity of hybridization spots per 
      nucleus were revealed between CaSki, SiHa and HeLa cells.(ABSTRACT TRUNCATED AT 
      250 WORDS)
FAU - Lizard, G
AU  - Lizard G
AD  - Centre Commun de Cytometrie en Flux, INSERM U80, Hopital E. Herriot, Lyon, 
      France.
FAU - Chignol, M C
AU  - Chignol MC
FAU - Souchier, C
AU  - Souchier C
FAU - Schmitt, D
AU  - Schmitt D
FAU - Chardonnet, Y
AU  - Chardonnet Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Histochemistry
JT  - Histochemistry
JID - 0411300
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Cell Nucleus/chemistry/virology
MH  - DNA, Viral/*analysis
MH  - Female
MH  - Genome, Viral
MH  - HeLa Cells
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Lasers
MH  - Microscopy/methods
MH  - Microscopy, Fluorescence
MH  - Papillomaviridae/*genetics
MH  - Papillomavirus Infections/genetics/virology
MH  - Sensitivity and Specificity
MH  - Tumor Virus Infections/genetics/virology
MH  - Uterine Neoplasms/virology
MH  - Virus Integration
EDAT- 1994/04/01 00:00
MHDA- 1994/04/01 00:01
CRDT- 1994/04/01 00:00
PHST- 1994/04/01 00:00 [pubmed]
PHST- 1994/04/01 00:01 [medline]
PHST- 1994/04/01 00:00 [entrez]
AID - 10.1007/BF00315918 [doi]
PST - ppublish
SO  - Histochemistry. 1994 Apr;101(4):303-10. doi: 10.1007/BF00315918.