PMID- 2478614
OWN - NLM
STAT- MEDLINE
DCOM- 19891127
LR  - 20170214
IS  - 0022-1554 (Print)
IS  - 0022-1554 (Linking)
VI  - 37
IP  - 11
DP  - 1989 Nov
TI  - Denaturation and condensation of intracellular nucleic acids monitored by 
      fluorescence depolarization of intercalating dyes in individual cells.
PG  - 1699-704
AB  - The intercalating binding of planar aromatic dye molecules to nucleic acids can 
      be analyzed using fluorescence depolarization measurements of the dye molecules 
      excited by linearly polarized light. In this study, we investigated the 
      conformational changes of the intracellular DNA-dye complex in single cells. Flow 
      cytometry, combined with a newly developed double-beam autocompensation 
      technique, permitted rapid high-precision fluorescence depolarization 
      measurements on a large number of individual cells. The dyes ethidium bromide 
      (EB), propidium iodide (PI), and acridine orange (AO) were used in this study. 
      Depending on the dye-to-phosphate ratio of the nuclear acid-dye complex, as well 
      as on the spatial dye structure itself, internal and external binding sites can 
      be monitored by fluorescence depolarization analysis. Both energy transfer and 
      rotation and vibration of the dye molecules cause depolarization of the 
      fluorescence emission. Differences in the concentration-dependent dye 
      fluorescence depolarization values between PI and EB on one side and AO on the 
      other side can be interpreted as a denaturation and condensation of 
      double-stranded DNA regions by AO. We further show that the fluorescence 
      polarization measurement technique can be used in an alternative way to monitor 
      thermal denaturation of cellular DNA.
FAU - Beisker, W
AU  - Beisker W
AD  - GSF Institut fur Biophysikalische Strahlenforschung, Neuherberg, Federal Republic 
      of Germany.
FAU - Eisert, W G
AU  - Eisert WG
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Histochem Cytochem
JT  - The journal of histochemistry and cytochemistry : official journal of the 
      Histochemistry Society
JID - 9815334
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Cell Line
MH  - Cricetinae
MH  - Cricetulus
MH  - DNA/*analysis
MH  - Flow Cytometry/methods
MH  - Fluorescence
MH  - Humans
MH  - *Nucleic Acid Conformation
MH  - *Nucleic Acid Denaturation
MH  - Staining and Labeling
EDAT- 1989/11/01 00:00
MHDA- 1989/11/01 00:01
CRDT- 1989/11/01 00:00
PHST- 1989/11/01 00:00 [pubmed]
PHST- 1989/11/01 00:01 [medline]
PHST- 1989/11/01 00:00 [entrez]
AID - 10.1177/37.11.2478614 [doi]
PST - ppublish
SO  - J Histochem Cytochem. 1989 Nov;37(11):1699-704. doi: 10.1177/37.11.2478614.