PMID- 2448084
OWN - NLM
STAT- MEDLINE
DCOM- 19880314
LR  - 20190828
IS  - 0271-3683 (Print)
IS  - 0271-3683 (Linking)
VI  - 6
IP  - 12
DP  - 1987 Dec
TI  - The evaluation of endothelial damage following corneal storage: a comparison of 
      staining methods and the value of scanning electron microscopy.
PG  - 1441-51
AB  - Staining techniques and scanning electron microscopy (SEM) were used to assess 
      damage to the endothelial cells following corneal storage. The aim was to 
      establish whether these assays are useful in the assessment of endothelial 
      integrity following corneal storage. Four groups ensured a range of normal and 
      damaged endothelial cells: 1) fresh; 2) stored for 7 days in a moist chamber; 3) 
      stored by the cryopreservation method of Capella and Kaufman; 4) damaged by rapid 
      freezing with a cryoprobe. Trypan blue (TB), nitroblue tetrazolium (NBT), 
      acridine orange (AO), fluorescein diacetate (FDA) and ethidium bromide (EB), were 
      used to stain the endothelial layer. SEM was carried out on duplicate samples. 
      Staining with NBT resulted in low cell counts due to loss of cells. There was no 
      significant difference between the extent of damage measured by TB, AO or FDA, 
      but it was shown that staining with FDA and EB can distinguish between damaged 
      and intact cells. Tissue stored for 7 days in a moist chamber had a reduced 
      number of intact cells compared to fresh tissue, and tissue stored by the Capella 
      and Kaufman technique gave a reduced number of intact cells compared to both 
      these control and storage groups. SEM showed surface perforation was 
      characteristic of injured cells, rather than complete disruption. FDA has a 
      theoretical advantage over the other stains, and should provide a more accurate 
      appraisal of defects of cell membrane integrity. For this reason, the use of FDA 
      with EB to stain the endothelium, with SEM carried out on duplicate samples, were 
      preferred as assays to use in the development of corneal storage.
FAU - Madden, P W
AU  - Madden PW
AD  - Department of Surgery, Cambridge University, UK.
LA  - eng
GR  - Wellcome Trust/United Kingdom
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Curr Eye Res
JT  - Current eye research
JID - 8104312
SB  - IM
MH  - Animals
MH  - Cell Count
MH  - Endothelium, Corneal/cytology/*ultrastructure
MH  - Evaluation Studies as Topic
MH  - Microscopy, Electron, Scanning/*standards
MH  - Rabbits
MH  - Staining and Labeling/*standards
MH  - *Tissue Preservation/methods
EDAT- 1987/12/01 00:00
MHDA- 1987/12/01 00:01
CRDT- 1987/12/01 00:00
PHST- 1987/12/01 00:00 [pubmed]
PHST- 1987/12/01 00:01 [medline]
PHST- 1987/12/01 00:00 [entrez]
AID - 10.3109/02713688709044508 [doi]
PST - ppublish
SO  - Curr Eye Res. 1987 Dec;6(12):1441-51. doi: 10.3109/02713688709044508.