PMID- 7537648
OWN - NLM
STAT- MEDLINE
DCOM- 19950608
LR  - 20131121
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 19
IP  - 3
DP  - 1995 Mar 1
TI  - Flow cytometric detection of micronuclei by combined staining of DNA and 
      membranes.
PG  - 201-8
AB  - A new staining method is presented for flow cytometric measurement of micronuclei 
      (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent 
      dyes in addition to DNA staining. Several combinations of fluorescent membrane 
      and DNA dyes were studied for a better discrimination of MN from debris in a 
      suspension of nuclei and micronuclei. For staining of membranes, the lipophilic 
      dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 
      1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium 
      bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral 
      properties, HO or EB combined with HEPn were not as suitable for the 
      discrimination of MN from debris as was HEPn in combination with PF. With HEPn in 
      combination with PF, however, additional noise was found at low fluorescence 
      intensities, probably due to free fluorescent dye molecules in the solution. The 
      optimal simultaneous staining of membranes and DNA was obtained using a 
      combination of DPH and EB. The induction of MN in Chinese hamster and mouse 
      NIH-3T3 cells by UV-B illumination was studied with this new staining technique. 
      UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster 
      cells were found to be more sensitive to these wavelengths. Illumination with 
      wavelengths above 360 nm did not induce MN in either cell line. The results 
      obtained from human lymphocytes using the combination of EB and DPH were 
      comparable to the results obtained with the combination of EB and HO.
FAU - Wessels, J M
AU  - Wessels JM
AD  - GSF-Forschungszentrum fur Umwelt und Gesundheit, Institut fur Biophysikalische 
      Strahlenforschung, Oberschleissheim, Germany.
FAU - Nusse, M
AU  - Nusse M
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Porphyrins)
RN  - 100572-96-1 (porphycene)
RN  - 1720-32-7 (Diphenylhexatriene)
RN  - 9007-49-2 (DNA)
RN  - CY3RNB3K4T (Proflavine)
RN  - EN464416SI (Ethidium)
RN  - LHQ7J5KV9B (Bisbenzimidazole)
SB  - IM
MH  - 3T3 Cells
MH  - Animals
MH  - Bisbenzimidazole
MH  - Cell Line
MH  - *Cell Membrane
MH  - Cricetinae
MH  - Cricetulus
MH  - DNA/*analysis
MH  - Diphenylhexatriene
MH  - Ethidium
MH  - Flow Cytometry/*methods
MH  - *Fluorescent Dyes
MH  - Humans
MH  - Lymphocytes
MH  - Mice
MH  - *Micronuclei, Chromosome-Defective/chemistry
MH  - Porphyrins
MH  - Proflavine
EDAT- 1995/03/01 00:00
MHDA- 1995/03/01 00:01
CRDT- 1995/03/01 00:00
PHST- 1995/03/01 00:00 [pubmed]
PHST- 1995/03/01 00:01 [medline]
PHST- 1995/03/01 00:00 [entrez]
AID - 10.1002/cyto.990190303 [doi]
PST - ppublish
SO  - Cytometry. 1995 Mar 1;19(3):201-8. doi: 10.1002/cyto.990190303.