Immunogold staining

Fig. 10.2. Immunogold staining of an ultra-thin section of an immature Acanthocheilonema viteae uterine mf with mAb 24-4. Note that A. viteae chitinase is present in the cuticle (arrowheads), but not on the surface. (Photograph W. Rudin.)...

Cubie, H.A., and Norval, M. (1989) Detection of human papilloma viruses in paraffin wax sections with biotinylated synthetic oligonucleotide probes and immunogold staining./. Clin. Pathol. 42, 988-991. 

Nielsen, M.H., Bastholm, L., Chatterjee, S., Koga, J., and Norrild, B. (1989) Simultaneous triple-immunogold staining of virus and host cell antigens with monoclonal antibodies of virus and host cell antigens in ultrathin cryosections. Elistochemistry 92, 89-93. 

Immunogold staining can be used successfully at the light microscopic level if the gold is silver-enhanced. Enhancing soluhons may be made up in the laboratory, but because of their instability and light sensitivity, the commercially available silver-enhancing kits are preferable. 

De Waele, M., De Mey, K., Moeremans, M., De Brabender, M., and Van Camp, B (1983) Immunogold staining for the light microscopic detection of leukocyte cell-surface antigens with monoclonal antibodies. J Histochem. Cytochem 31, 938-944. 

Another recent study also supports the mediation role of EDTA in unmasking antigens (Rocken and Roessner, 1999). In this study thin sections of aldehyde-fixed, Epon-embedded human biopsy tissues were treated with 1 mM EDTA, using a heated water bath. This combined treatment resulted in excellent immunogold staining of amyloid (Fig. 6.4). 

IL-5 BAL cells of asthmatics In situ hybridization EM immunogold staining Broide et at. (1992b)... 

HES Eosinophilic cystitis in situ hybridization EM immunogold-staining Immunohistochemistry Dubucquoi et al. (1994)... 

Fig. 4. Disruption of the synapsin-dependent vesicle pool by presynaptic microinjection of synapsin antibodies in the lamprey reticulospinal synapse. (A) Electron micrograph of a control synapse. (B) A synapse in an axon injected with synapsin antibodies. The axon was lightly stimulated (1 Hz for 12 min) and allowed to rest for 90 min before fixation. Note that a narrow rim of vesicles remains in the antihody-injected synapse. (O Immunogold staining of a reticulospinal synapse with synapsin antibodies. Note that the vesicles adjacent to the presynaptic membrane are almost devoid of gold particles. (D) Visualization of the filamentous cytomatrix that overlaps with the synaptic vesicle pool that remains after perturbation of synapsins. The electron micrograph shows a synapse in a normal axon (i.e. no microinjection or stimulation had been performed) stained with phosphotungstic acid (Gustafsson et al., 1996). The filamentous eytomatrix (arrowheads) at the presynaptic membrane is visible, but not the synaptic vesicle cluster. Designations as in Fig. 1. Scale bar, 0.2 p.m. Reprinted from Brodin et al. (1995) Eur J Neurosci 9 2503-2511, with permission.

As with the animal monomeric and heterotrimeric G-proteins, the proteins identified appeared to show differential expression within the plant tissues. Northern blot analysis showed that transcripts encoding GPAl were most abundant in vegetative tissue and least abundant in floral and apical meristems [89], whilst immunogold staining supported this and also showed that GPAl was strongly expressed in vascular tissue [153]. 

Transmission electron microscopy analysis revealed a marked expansion of the endomysial space in thyroid eye disease extraocular muscle biopsies compared with that in control biopsies (69). An increased number of collagen fibers with hyaluronan were detected by immunogold staining, although the serum levels of hyaluronan and urinary glycosaminoglycans were not found to be sensitive indicators for the presence of these molecules within the extraocular muscles (69). Imai et al. further confirmed that the local accumulation of glycosaminoglycans in thyroid eye disease was not associated with the serum hyaluronan concentration (70). 

Appendix I Preembedding Immunogold Staining Sample Protocol... 

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