Document Detail


Microwave-stimulated incubation in immunoelectron microscopy: a quantitative study.
MedLine Citation:
PMID:  3220797     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Microwave irradiation has been applied to reduce the immunogold staining time of ultrathin sections of Lowicryl embedded specimens. Labelling has been stimulated by microwave irradiation during incubation with 10 nm colloidal gold particles coated with either goat anti-mouse antibodies (GaM-gold) or goat anti-rabbit antibodies (GaR-gold) and has been compared with control incubations. Quantification has been performed on cytoplasmic membranes or lysosomes labelled with a primary antibody. Counting the gold particles over specific and non-specific sites in electron micrographs and electron microscopic images by IBAS 2000 revealed that irradiation of 25 microliters droplets both at 80 W and 150 W resulted in an accelerated immunogold labelling, while the non-specific background levels were not increased. A plateau level in immunogold labelling intensity was reached after 25 min incubation under microwave irradiation at 150 W as compared to 120 min incubation without microwaves. No improvement in localization sharpness of immunogold labelling on membranes was achieved by microwave irradiation. The microwave-mediated acceleration of immunogold staining may be considered as an example of a staining method with a restricted thermal action on microvolumes as indicated by direct temperature measurements using a fibre-optic thermometer.
Authors:
P E Zondervan; A De Jong; C W Sorber; L P Kok; W C De Bruijn; T H Van der Kwast
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Histochemical journal     Volume:  20     ISSN:  0018-2214     ISO Abbreviation:  Histochem. J.     Publication Date:    1988 Jun-Jul
Date Detail:
Created Date:  1989-03-17     Completed Date:  1989-03-17     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0163161     Medline TA:  Histochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  359-64     Citation Subset:  IM    
Affiliation:
Department of Pathology, Erasmus University, Rotterdam, Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Antigens, Surface / analysis
Gold
Humans
Immunohistochemistry / methods
Lysosomes / ultrastructure
Male
Mesothelioma / pathology,  ultrastructure*
Microscopy, Electron / methods*
Microwaves*
Prostate / ultrastructure*
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, Surface; 7440-57-5/Gold

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


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