PMID- 3743111
OWN - NLM
STAT- MEDLINE
DCOM- 19861016
LR  - 20190829
IS  - 0271-3683 (Print)
IS  - 0271-3683 (Linking)
VI  - 5
IP  - 7
DP  - 1986 Jul
TI  - Supravital fluorescent staining of the corneal endothelium with acridine orange 
      and ethidium bromide.
PG  - 485-94
AB  - We investigated the combined fluorescent dyes acridine orange (AO) and ethidium 
      bromide (EB) for corneal endothelial evaluation. Both dyes intercalate with DNA 
      and RNA and are mutagenic at high concentrations. Optimum differential staining 
      was obtained after 5 minutes exposure to 1 microgram/ml of each dye. Electron 
      microscopy confirmed that the dye combination simultaneously identified both 
      viable and nonviable cells. Swelling of perfused dog corneas for 3 hours after 
      exposure to both dyes for 5 minutes was not increased. Exposure to visible light 
      from a fluorescence microscope for 2-5 minutes caused increased corneal swelling 
      and abnormal intercellular junctions by electron microscopy in paired dog corneas 
      stained for 5 minutes with either AO or EB. The addition of 10% calf serum to the 
      staining and observation media decreased the toxic effect of AO/EB and light seen 
      by specular microscopy and by electron microscopy. Human corneas exposed to AO 
      and EB were not mutagenic in the Salmonella/microsome assay. These results 
      indicate that AO and EB, 1 microgram/ml, provide a rapid identification of both 
      viable and nonviable corneal endothelial cells. When the corneas are exposed to 
      light, however, the dyes are toxic; this toxicity is decreased by the presence of 
      10% calf serum in the staining and observation media.
FAU - Kolb, M J
AU  - Kolb MJ
FAU - Bourne, W M
AU  - Bourne WM
LA  - eng
GR  - EY 02037/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Curr Eye Res
JT  - Current eye research
JID - 8104312
RN  - EN464416SI (Ethidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - *Acridine Orange
MH  - Animals
MH  - Cell Survival
MH  - Cornea/cytology/*physiology/ultrastructure
MH  - Dogs
MH  - Endothelium/cytology/physiology/ultrastructure
MH  - *Ethidium
MH  - Female
MH  - Humans
MH  - Male
MH  - Microscopy, Electron
MH  - Microscopy, Fluorescence
MH  - Mutagenicity Tests
MH  - Organ Culture Techniques
MH  - Rabbits
MH  - Time Factors
EDAT- 1986/07/01 00:00
MHDA- 1986/07/01 00:01
CRDT- 1986/07/01 00:00
PHST- 1986/07/01 00:00 [pubmed]
PHST- 1986/07/01 00:01 [medline]
PHST- 1986/07/01 00:00 [entrez]
AID - 10.3109/02713688608996370 [doi]
PST - ppublish
SO  - Curr Eye Res. 1986 Jul;5(7):485-94. doi: 10.3109/02713688608996370.