Electron microscopy immunogold labeling

Staining of consecutive sections with histochemical reagents for protein and antibodies against myrosinase showed that myrosin cells actually contain myrosinase (Fig. 4.2D,E).18,43,44 By electron microscopy immunogold labelling studies, the enzyme was localized to the interior of the vacuoles of the myrosin cells (Fig. 4.2F).18,40,41 It has also been shown by several in situ hybridization experiments that myrosinase transcripts are located in these cells (Fig. 4.2G),6 demonstrating that the cellular localization of myrosinase is due to transcriptional regulation and not to a transport process. No evidence for transport of myrosinase currently exists. These methods for detection and identification of myrosin cells and myrosinase expression are illustrated (Fig. 4.2). For a historical overview of myrosinase localization studies, see Bones and Rossiter.45... 

Fig. 10. Intercellular junction zones of carrot cells grown in suspension have been observed in electron microscopy after immunogold labeling with the 2F4 antibody, (a) no treatment of the sections prior to labeling the gold particles are restricted to the center of the junction zones (b) enzymatic (pectin methyl esterase) deesterification of the E.M. grids before labeling the deesterified pectins present in the primary walls now bind the probe. Scale bars = 1 pm.

NPQ (Rakhimberdieva et al. 2004) exactly matches the absorption spectrum of the carotenoid, 3 -hydrox yech i nenone (Polivka et al. 2005) in the OCP. The OCP is now known to be specifically involved in the phycobilisome-associated NPQ and not in other mechanisms affecting the levels of fluorescence such as state transitions or D1 damage (Wilson et al. 2006). Studies by immunogold labeling and electron microscopy showed that most of the OCP is present in the interthylakoid cytoplasmic region, on the phycobilisome side of the membrane, Figure 1.2 (Wilson et al. 2006). The existence of an interaction between the OCP and the phycobilisomes and thylakoids was supported by the co-isolation of the OCP with the phycobilisome-associated membrane fraction (Wilson et al. 2006, 2007). 

Hyatt AD. Immunogold labeling techniques, in Electron Microscopy in Biology— A Practical Approach (Harris JR, ed.), IRL Press, Oxford, UK, 1991, pp. 59-81. 

Vandenbosch KA. Immunogold labeling, in Electron Microscopy of Plant Cells (Hall JL, Hawes C, eds.), Academic Press, San Diego, CA, 1991, pp. 181-218. 

Birrell, G. B., Hedberg, K. K., and Griffith, P. H. (1987) Pitfalls of immunogold labeling analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy. J. Histochem. Cytochem. 35, 843-853. 

Transmission electron microscopy of immunogold labelled sections has shown that the extracellular lignin-degrading enzymes lignin-peroxidase and laccase were localized within the cell wall and mucilage of the hyphae of C. versicolor. Laccase was present in the cell wall layer whereas lignin-... 

Fig. 4 Transmission electron microscopy of a longitudinal section of the posterior end of a Cryptosporidium parvum sporozoite showing immunogold localization of pyruvate NADP+ oxidoreductase (CpPNO). The mitochondrion-like organelle ( ) is posterior to the nucleus, and lies between the nucleus and the CB. It is labeled by mitochondrion-specific 15-nm gold anti- particles. Small -nm gold goat anti-CpPFO particles (arrows) show the localization of CpPNO. There are no 6-nm particles localized within the mitochondrion-like organelle (reprinted from Fig. 12 of Ctrnacta et al. 2006 with permission of the publishers)...

Timms BG. (1989) Postembedding immunogold labeling for electron microscopy using "LR White" resin. Am. J. Anat. 175, 267-75. 

Ohno M, Takemura G, Ohno A, Misao J, Hayakawa Y, Minatoguchi S, et al. Apoptotic myocytes in infarct area in rabbit hearts may be oncotic myocytes with DNA fragmentation Analysis by immunogold electron microscopy combined with in situ nick end-labeling. Circulation 1998 98 1422-1430. 

Transmission electron microscopy analyses of ultrathin sections have indicated that there are several vacuoles in myrosin cells of embryos. These vacuoles contain myrosinase, as revealed by immunogold labelling studies, and have been referred to as myrosin grains.40, 1 Recently, confocal laser scanning immuno-microscopy analysis with the anti-myrosinase monoclonal antibody 3D7 has been... 

Ultrathin sections for electron microscopy (thickness approximately 100 nm) must be able to withstand the electron beam and the vacuum in the microscope. For this, it is first necessary to stabilize the ultrastructure of the fresh tissue by fixation, then to dehydrate it with an organic solvent, and finally to embed it in a resin. The resulting hard block can be cut into ultrathin sections, which are then mounted on a grid and stained. For immunogold labeling, free-aldehyde groups and nonspecific binding sites... 

Hillbrick, G.C., McMahon, D.J., and McManus, W.R. (1999) Microstmcture of indirectly and directly heated ultra-high-temperature (UHT) processed milk examined using transmission electron microscopy and immunogold labelling. Lebensmiti. Wissen. Technol. 32, 486 94. 

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