Single and Double Colloidal Gold Labeling in Postembedding Immunoelectron Microscopy

2004 ◽  
pp. 161-170
Author(s):  
Nicoletta Zini ◽  
Liliana Solimando ◽  
Caterina Cinti ◽  
Nadir Mario Maraldi
Keyword(s):  
Immunoelectron Microscopy ◽  
Colloidal Gold ◽  
Postembedding Immunoelectron Microscopy

scholarly journals Microfibril-associated glycoprotein-1 (MAGP-1) is specifically located on the beads of the beaded-filament structure for fibrillin-containing microfibrils as visualized by the rotary shadowing technique.

1996 ◽  
Vol 44 (12) ◽  
pp. 1389-1397 ◽  
Author(s):  
M Henderson ◽  
R Polewski ◽  
J C Fanning ◽  
M A Gibson
Keyword(s):  
Monoclonal Antibody ◽  
Immunoelectron Microscopy ◽  
Narrow Band ◽  
Immunogold Labeling ◽  
Structural Role ◽  
Filament Structure ◽  
Fibrillin 1 ◽  
Postembedding Immunoelectron Microscopy ◽  
Microscopic Techniques ◽  
Rotary Shadowing

This study used immunoelectron microscopic techniques to define the ultrastructural location of MAGP-1 on the fibrillin-containing microfibrils of the ocular zonule. A specific anti-MAGP-1 monoclonal antibody (MAb), 11B, was produced that did not crossreact with fibrillin-1 or other microfibrillar proteins. MAb 11B was shown by immunofluorescence to localize intensely to zonular tissue. Postembedding immunoelectron microscopy showed that MAGP-1 was associated with microfibrils throughout the zonule, with the exception of a narrow band of microfibrils at the junction with the lens capsule. With preembedding labeling, the anti-MAGP-1 MAb was found to localize in a crossbanding pattern, at intervals of about 50 nm, to microfibrils throughout the zonule and along bundles of microfibrils in surrounding vitreous tissue. Rotary shadowing of isolated microfibrils showed a "beads on a string" morphology with a periodicity of about 50 nm. With immunogold labeling, the anti-MAGP-1 antibody specifically localized on the beads in a symmetrical manner. Occasionally two gold partides were attached to the same bead, suggesting that multiple MAGP-1 molecules were present in the structure. The results indicate that MAGP-1 is intimately and regularly associated with the bead regions of fibrillin-containing microfibrils. The findings are consistent with a major structural role for MAGP-1 in microfibril biology.

Immunoelectron microscopy provides a road-map for antigen processing and presentation

1990 ◽  
Vol 48 (3) ◽  
pp. 40-40
Author(s):  
Lynne Guagliardi ◽  
Debra Crumrine ◽  
Frances Brodsky
Keyword(s):  
Immunoelectron Microscopy ◽  
Colloidal Gold ◽  
Antigen Processing ◽  
Uranyl Acetate ◽  
Endocytic Pathway ◽  
Road Map ◽  
Cacodylate Buffer ◽  
Intracellular Compartments ◽  
Antigen Processing And Presentation ◽  
Human B Cell

Immunoelectron microscopy was used to identify the intracellular routes traveled by molecules involved in antigen processing and presentation by human B cell lines. To mimic the uptake of multivalent antigen, anti-immunoglobulin coupled to 15-nm colloidal gold beads was used to cross-link surface immunoglobulin and induce its internalization at 37°C. At various timepoints following internalization, cells were fixed in cacodylate buffer with 2% paraformaldehyde, postfixed briefly with OsO4 ferricyanide, and en-block stained with aqueous uranyl acetate. After brief dehydrations in 50% and 70% ethanol, cells were gradually infiltrated with L.R. White Resin (medium hardness) and polymerized at 50°C overnight. 60-nm sections were labelled with monoclonal antibodies and polyclonal antisera adsorbed to 2- or 5-nm colloidal gold to characterize the intracellular compartments containing immunoglobulin with respect to their pH, enzyme content, the presence of Class II histocompatibility molecules, the invariant chain, and markers for the endocytic pathway. Using 2 sizes of gold probes, we simultaneously mapped the distribution of 2 different molecules in the same immunoglobulin-containing compartment.

Lateral mobility of membrane-bound antibodies on the surface of Acholeplasma laidlawii: evidence for virus-induced cell fusion in a procaryote

1982 ◽  
Vol 152 (1) ◽  
pp. 471-478
Author(s):  
K Haberer ◽  
D Frösch
Keyword(s):  
Cell Fusion ◽  
Immunoelectron Microscopy ◽  
Colloidal Gold ◽  
Dynamic Processes ◽  
Cell Populations ◽  
Temperature Dependent ◽  
Lateral Mobility ◽  
Acholeplasma Laidlawii ◽  
Membrane Bound ◽  
First Time

Dynamic processes on the membrane of the procaryotic cell Acholeplasma laidlawii have been studied by means of immunoelectron microscopy. Colloidal gold-labeled anti-A. laidlawii antibodies were used as electron-dense markers. This method allowed the demonstration of temperature-dependent lateral mobility of membrane-bound immunoglobulins. By using two different sizes of gold grains to differentiate cells from two different cell populations, virus-induced fusion of procaryotic cells could be shown for the first time.

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