Summary
The application of an automatic image analyzer (TAS, Leitz, Wetzlar) for determination of labeling density in protein A-gold labeled sections is described. Electron micrographs of rat liver labeled with 12 nm gold particles for peroxisomal enzymes are placed on the macrounit of TAS and the images of peroxisomes on TAS-monitor are contoured with a light pen. The instrument measures the surface of the contoured areas. Based on their gray level, the gold particles over the peroxisome are detected automatically and counted and the labeling density for each peroxisome is calculated.
References
Bendayan M (1984) Protein A-gold electron microscopic immunocytochemistry: Methods, applications and limitations. J Electron Microsc Tech 1:243–270
Bendayan M, Reddy JK (1982) Immunocytochemical localization of catalase and heat-labile enoyl-CoA hydratase in the livers of normal and peroxisome proliferator-treated rats. Lab Invest 47:364–369
Bendayan M, Roth J, Perrelet A, Orci L (1980) Quantitative immunocytochemical localization of pancreatic secretory proteins in subcellular compartments of the rat acinar cell. J Histochem Cytochem 28:149–160
Fahimi HD, Reinicke A, Sujatta M, Yokota S, Özel M, Hartig F, Stegmeier K (1982) The short- and long-term effects of bezafibrate in the rat. Ann NY Acad Sci 386:111–135
Nanney LB, McKanna JA, Stoscheck CM, Carpenter G, King LE (1984) Visualization of epidermal growth factor receptors in human epidermis. J Invest Dermatol 82:165–169
Ratcliffe A, Fryer PR, Hardingham TE (1984) The distribution of aggregating proteoglycans in articular cartilage. J Histochem Cytochem 32:193–201
Ravazzola M, Perrelet A, Roth J, Orci L (1981) Insulin immunoreactive sites demonstraded in the Golgi apparatus of pancreatic B cells. Proc Natl Acad Sci USA 78:5661–5664
Roth J (1982) The protein A-gold technique — a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, Vol. 1, Academic Press, New York, pp 108–133
Thorens B, Roth J, Norman AW, Perrelet A, Orci L (1982) Immunocytochemical localization of the vitamin D-dependent calcium binding protein in chick duodenum. J Cell Biol 94:115–122
Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30:23–38
Yokota S, Oda T (1983) Immunoelectron microscopic localization of serine: Pyruvate aminotransferase in rat eosinophil leukocytes. Histochemistry 78:417–424
Yokota S, Deimann W, Hashimoto T, Fahimi MD (1983a) Immunocytochemical localization of two peroxisomal enzymes of lipid β-oxidation in specific granules of rat cosinophils. Histochemistry 78:425–433
Yokota S, Hashimoto T, Fahimi HD (1983b) Fatty acid β-oxidation enzymes in peroxisomes: Comparison of localization by pre- and postembedding immunocytochemical techniques. Fed Proc 42:1303 (Abstract)
Yokota S, Tsuji H, Kato K (1984a) Localization of lysosomal and peroxisomal enzymes in the specific granules of rat intestinal eosinphil leukocytes revealed by immunoelectron microscopic techniques. J Histochem Cytochem 32:267–274
Yokota S, Deimann W, Hashimoto T, Fahimi HD (1984b) Specific granules of rat eosinophils contain peroxisomal acyl-CoA oxidase: possible involvement in production of H2O2. Histochem J 16:573–577
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Beier, K., Fahimi, H.D. Automatic determination of labeling density in protein A-gold immunocytochemical preparations using an image analyzer. Histochemistry 82, 99–100 (1985). https://doi.org/10.1007/BF00502096
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DOI: https://doi.org/10.1007/BF00502096