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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
6
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pubmed:dateCreated |
1997-3-4
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pubmed:abstractText |
The purpose of this study was to modify the gold chloride procedure for studies of total innervation in corneal whole mounts to provide a decrease in nonspecific background staining and to eliminate the progressively deteriorating stain quality of standard gold chloride techniques. Modifications included use of cryoprotective agents, mechanical removal of Descemet's membrane-endothelium complex prior to fixation, treatment with alpha amylase, and halting the reduction of gold chloride to metallic gold using Kodak rapid fixer with hardener. Rat corneas were stored at -70 C in O.C.T. compound. The Descemet's membrane-endothelium complex was removed after thawing, and corneas were fixed in 4% NaPO4-buffered paraformaldehyde with 8% sucrose. Fixed corneas were incubated in NaPO4-buffered saline containing alpha amylase, placed in 100% lemon juice, then in 1% gold chloride solution, transferred to glacial acidic acid, placed in rapid fixer, rinsed in NaPO4-buffered saline, and dehydrated in graded alcohols. Flat mounts of whole corneas were examined using contralateral corneas as controls. Freezing corneas in O.C.T. compound, removal of the Descemet's membrane-endothelium complex, and treatment with alpha amylase reduced non-specific background staining compared to controls. Treatment with Kodak rapid fixer prevented the deterioration of staining quality for at least 8 months. These improvements allow the gold chloride technique to be used with immunohistochemical procedures where the reaction products would be obscured by background staining.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
1052-0295
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
70
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
277-84
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:9044655-Animals,
pubmed-meshheading:9044655-Cornea,
pubmed-meshheading:9044655-Cryoprotective Agents,
pubmed-meshheading:9044655-Eye Enucleation,
pubmed-meshheading:9044655-Gold Compounds,
pubmed-meshheading:9044655-Histocytochemistry,
pubmed-meshheading:9044655-Plastic Embedding,
pubmed-meshheading:9044655-Rats,
pubmed-meshheading:9044655-Rats, Sprague-Dawley,
pubmed-meshheading:9044655-Staining and Labeling,
pubmed-meshheading:9044655-Tissue Fixation,
pubmed-meshheading:9044655-alpha-Amylases
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pubmed:year |
1995
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pubmed:articleTitle |
Improved gold chloride procedure for nerve staining in whole mounts of rat corneas.
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pubmed:affiliation |
Section on Pathophysiology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892-2740, USA.
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pubmed:publicationType |
Journal Article
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