20:
121:
found in low densities, such as some cell surface antigens. As the resolution of scanning electron microscopy (SEM) increased, so too did the need for nanoparticle-sized labels such as immunogold. In 1975, Horisberger and coworkers successfully visualised gold nanoparticles with a diameter of less
141:
The prepared sample is then incubated with a specific antibody designed to bind the molecule of interest. Next, a secondary antibody which has gold particles attached is added, and it binds to the primary antibody. Gold can also be attached to
677:
Van Laere O, De Wael L, De Mey J (1985). "Immuno gold staining (IGS) and immuno gold silver staining (IGSS) for the identification of the plant pathogenic bacterium
Erwinia amylovora (Burrill) Winslow et al".
192:. The silver enhancement increases the particle size, also making scanning electron microscopy possible. In order to produce the silver-enhanced gold particles, colloidal gold particles are placed in an
255:
used to embed samples for imaging. Thus, only accessible molecules can be targeted and visualized. Labeling prior to embedding the sample can reduce the negative impact of this limitation.
100:
Immunogold labeling can introduce artifacts, as the gold particles reside some distance from the labelled object and very thin sectioning is required during sample preparation.
225:
An inherent limitation to the immunogold technique is that the gold particle is around 15-30 nm away from the site to which the primary antibody is bound (when using a
244:
may appear as a 'spike' depending on which plane the sectioning occurred. To overcome this limitation serial sections can be taken, which can then be compiled into a
229:
labeling strategy). The precise location of the targeted molecule can therefore not be accurately calculated. Gold particles can be created with a diameter of 1
236:
Thin sections are required for immunogold labeling and these can produce misleading images; a thin slice of a cell component may not give an accurate view of its
208:
site and silver is deposited onto the particle. An example of the application of silver-enhanced immunogold labeling (IGSS) was in the identification of the
172:
by using two different-sized gold particles. An extension of this method used three different sized gold particles to track the localisation of regulatory
385:
188:
Although immunogold labeling is typically used for transmission electron microscopy, when the gold is 'silver-enhanced' it can be seen using
160:
The electron-dense gold particle can now be seen under an electron microscope as a black dot, indirectly labeling the molecule of interest.
19:
728:
233:(or lower) but another limitation is then realized—at these sizes the gold label becomes hard to distinguish from tissue structure.
226:
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63:
340:
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51:
97:. The labeling technique can be adapted to distinguish multiple objects by using differently-sized gold particles.
23:
Two images produced using immunogold labeling and transmission electron microscopy: (A) Gold particles are marking
559:"Coloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques"
600:"Double immunogold staining method for the simultaneous ultrastructural localization of regulatory peptides"
245:
117:. It was first applied in transmission electron microscopy (TEM) and was especially useful in highlighting
189:
168:
Immunogold labeling can be used to visualize more than one target simultaneously. This can be achieved in
94:
28:
718:
639:
Bendayan M (January 1982). "Double immunocytochemical labeling applying the protein A-gold technique".
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197:
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separately, the immunogold particles attached to both sides can then be viewed simultaneously.
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Hermann R, Walther P, Müller M (1996). "Immunogold labeling in scanning electron microscopy".
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483:"Ultrastructural localization of intracellular antigens by the use of protein A-gold complex"
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Faulk WP, Taylor GM (November 1971). "An immunocolloid method for the electron microscope".
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75:
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176:. A more complex method of multi-site labeling involves labeling opposite sides of an
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A further limitation is that antibodies and gold particles cannot penetrate the
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59:
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147:
143:
131:
543:
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Porter, K; Blum, J (1953). "A study in
Microtomy for Electron Microscopy".
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Immunogold labeling was first used in 1971 by Faulk and Taylor to identify
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285:"The functional organization of mitochondrial genomes in human cells"
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than 30 nm and this soon became an established SEM technique.
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Tapia FJ, Varndell IM, Probert L, De Mey J, Polak JM (July 1983).
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instead of a secondary antibody, as these proteins bind mammalian
24:
18:
85:
First used in 1971, immunogold labeling has been applied to both
201:
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First, a thin section of the sample is cut, often using a
31:(B) mtDNA marked with gold particles after extraction.
341:"Immunogold Labeling in Scanning Electron Microscopy"
50:. This staining technique is an equivalent of the
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670:
58:particles are most often attached to secondary
16:Staining technique used in electron microscopy
8:
481:Roth J, Bendayan M, Orci L (December 1978).
82:scatter to give high contrast 'dark spots'.
380:(5th ed.). New York: Garland Science.
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283:Iborra FJ, Kimura H, Cook PR (2004).
74:component. Gold is used for its high
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376:Alberts, Bruce; et al. (2008).
46:) is a staining technique used in
14:
227:primary and secondary antibodies
87:transmission electron microscopy
557:Roth J, Binder M (March 1978).
445:Histochemistry and Cell Biology
204:. Gold particles then act as a
184:Uses in brightfield microscopy
62:which are in turn attached to
1:
378:Molecular biology of the cell
54:technique for visible light.
415:10.1016/0019-2791(71)90496-4
91:scanning electron microscopy
66:designed to bind a specific
238:three-dimensional structure
52:indirect immunofluorescence
745:
729:Electron microscopy stains
134:. Various other stages of
164:Labeling multiple objects
157:in a non-specific way.
641:J. Histochem. Cytochem
604:J. Histochem. Cytochem
563:J. Histochem. Cytochem
487:J. Histochem. Cytochem
190:brightfield microscopy
95:brightfield microscopy
32:
536:10.1002/ar.1091170403
524:The Anatomical Record
303:10.1186/1741-7007-2-9
138:may then take place.
22:
653:10.1177/30.1.6172469
617:10.1177/31.7.6189888
500:10.1177/26.12.366014
265:Immunohistochemistry
576:10.1177/26.3.632554
170:electron microscopy
48:electron microscopy
40:immunogold staining
36:Immunogold labeling
692:10.1007/BF00509198
457:10.1007/BF02473200
200:containing silver
136:sample preparation
64:primary antibodies
33:
387:978-0-8153-4106-2
246:three-dimensional
240:. For example, a
214:Erwinia amylovora
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343:. Archived from
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78:which increases
76:electron density
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493:(12): 1074–81.
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403:Immunochemistry
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680:Histochemistry
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530:(4): 685–710.
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409:(11): 1081–3.
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178:antigenic site
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56:Colloidal gold
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610:(7): 977–81.
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719:Biochemistry
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349:. Retrieved
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29:mitochondria
647:(1): 81–5.
242:microtubule
221:Limitations
724:Microscopy
713:Categories
351:2010-07-08
271:References
206:nucleation
196:enhancing
155:Fc regions
111:Salmonella
60:antibodies
290:BMC Biol.
148:protein G
144:protein A
132:microtome
126:Technique
70:or other
27:near the
544:13124776
322:15157274
259:See also
210:pathogen
198:solution
174:peptides
119:proteins
115:antigens
80:electron
700:2416717
661:6172469
626:6189888
465:8858365
423:4110101
248:image.
104:History
68:antigen
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585:632554
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509:366014
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313:425603
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194:acidic
296:: 9.
253:resin
25:mtDNA
696:PMID
657:PMID
622:PMID
581:PMID
540:PMID
505:PMID
461:PMID
419:PMID
382:ISBN
318:PMID
202:ions
89:and
72:cell
688:doi
649:doi
612:doi
571:doi
532:doi
528:117
495:doi
453:doi
449:106
411:doi
308:PMC
298:doi
152:IgG
146:or
44:IGS
38:or
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Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
