Document Detail


Parameters affecting imaging of the horseradish-peroxidase-diaminobenzidine reaction product in the confocal scanning laser microscope.
MedLine Citation:
PMID:  2084264     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Neurons intracellularly filled with biocytin and labelled with nickel-intensified diaminobenzidine (DAB/Ni) can be imaged on the confocal scanning laser microscope in order to obtain three-dimensional and optical section images of neurons. Intensification of the DAB reaction product with nickel was found to be crucial for obtaining a workable signal level. On the other hand, the high absorption of light by the reaction product severely attenuated the detection of structures lying directly underneath, and the intensity of the unattenuated laser used for imaging faded or damaged DAB/Ni reaction product. We have determined that reduction of the laser intensity combined with the use of proper objective lenses and non-laser-based imaging for preliminary adjustments of the specimen all work to reduce or eliminate damage, and also improve the image. These items must be kept in mind when imaging and analysing DAB-labelled structures in the laser-based confocal microscope.
Authors:
J S Deitch; K L Smith; J W Swann; J N Turner
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of microscopy     Volume:  160     ISSN:  0022-2720     ISO Abbreviation:  J Microsc     Publication Date:  1990 Dec 
Date Detail:
Created Date:  1991-05-15     Completed Date:  1991-05-15     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  265-78     Citation Subset:  IM    
Affiliation:
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany.
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MeSH Terms
Descriptor/Qualifier:
3,3'-Diaminobenzidine*
Animals
Dendrites / ultrastructure
Hippocampus
Horseradish Peroxidase*
Lasers*
Microscopy / methods*
Neurons / ultrastructure*
Rats
Spectrophotometry
Grant Support
ID/Acronym/Agency:
NS18309/NS/NINDS NIH HHS; RR01219/RR/NCRR NIH HHS; RR02984/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
91-95-2/3,3'-Diaminobenzidine; EC 1.11.1.-/Horseradish Peroxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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