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Light microscopical detection of leukocyte cell surface antigens with a one-nanometer gold probe

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Summary

The potential of ultrasmall gold particles for the light microscopical detection of leukocyte cell surface differentiation antigens was investigated. Suspensions and cytocentrifuge preparations of peripheral blood leukocytes were first incubated with monoclonal antibodies and then with goat antimouse antibodies coupled to colloidal gold particles of 1-nanometer diameter. Cytocentrifuge preparations were made from the cell suspensions. Silver enhancement was performed on all preparations. Then they were counterstained with May-Grünwald Giemsa and examined in light microscopy. The immunostaining appeared as fine dark granules on the surface membrane of the cells. Labeling conditions were determined which gave a dense specific immunostaining and a low background. High dilutions of the ultrasmall gold probe could be used to detect all antigen expressing cells in the samples. The labeling efficiency of the IGSS method with the 1 nanometer probe was comparable to that described earlier for 5 nanometer gold particles. Lymphocyte subsets enumerated with this method in normal peripheral blood were similar to those found with immunofluorescence microscopy. We concluded that one nanometer probes do not offer a major advantage in comparison with 5 nanometer probes for the study of cell surface antigens.

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References

  • Baschong W, Lucocq JM, Roth J (1985) Thiocyanate gold: small (2–3 nm) colloidal gold for affinity cytochemical labeling in electron microscopy. Histochemistry 83:409–411

    Google Scholar 

  • Behnke O, Ammitzboll T, Jessen H, Klokker M, Nilausen K, Transum-Jensen J, Olsson L (1986) Nonspecific binding of proteinstabilized gold sols as a source of error in immunocytochemistry. Eur J Cell Biol 41:326–338

    Google Scholar 

  • Birrel GB, Hedberg KK, Hayes-Griffith O (1987) Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy and photoelectron microscopy. J Histochem Cytochem 35:843–853

    Google Scholar 

  • Crockard A, Catovsky D (1983) Cytochemistry of normal lymphocyte subsets defined by monoclonal antibodies and immunocolloidal gold. Scand J Haematol 30:433–443

    Google Scholar 

  • Danscher G (1981) Localization of gold in biological tissue. A photochemical method for light and electron microscopy. Histochemistry 71:81–88

    Google Scholar 

  • Danscher G (1983) A silver method for counterstaining plastic embedded tissue. Stain Technol 58:365–372

    Google Scholar 

  • Danscher G, Nörgaard R (1983) Light microscopic visualisation of colloidal gold in resin-embedded tissue. J Histochem Cytochem 31:1394–1398

    Google Scholar 

  • De Mey J (1983) Colloidal gold probes in immunocytochemistry. In: Polak J, Van Noordende S (eds) Immunocytochemistry: applications in pathology and biology. Wright and Sons, London, pp 82–112

    Google Scholar 

  • De Mey J (1984) Colloidal gold as marker and tracer in light-and electron microscopy. Electron Microsc Soc Am Bull 14:54

    Google Scholar 

  • De Mey J, Moeremans M, Geuens G, Nuydens R, De Brabander M (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immunogold staining) method. Cell Biol Int Rep 5:889–899

    Google Scholar 

  • De Waele M (1984) Hematological electron immunocytochemistry. Detection of cell surface antigens with monoclonal antibodies. In: Polak J, Varndell I (eds) Immunolabeling for electron microscopy. Elsevier, Amsterdam, pp 267–288

    Google Scholar 

  • De Waele M (1989) Silver-enhanced colloidal gold for the detection of leukocyte cell surface antigens in darkfield and epipolarization microscopy. In: Hayat MA (ed) Colloidal gold: principles, methods and applications, vol 2. Academic Press, San Diego, Calif., USA, pp 443–467

    Google Scholar 

  • De Waele M, De Mey J, Moeremans M, De Brabander M, Van Camp B (1983) Immunogold staining method for the light microscopic detection of leukocyte cell surface antigens with monoclonal antibodies. Its application to the enumeration of lymphocyte subpopulations. J Histochem Cytochem 31:376–381

    Google Scholar 

  • De Waele M, De Mey J, Moeremans M, Broodtaerts L, Smet L, Van Camp B (1982) Colloidal gold as marker for the light microscope detection of leukocyte cell surface antigens with monoclonal antibodies. J Clin Immunol 2:24–31

    Google Scholar 

  • De Waele M, De Mey J, Renmans W, Labeur C, Jochmans K, Van Camp B (1986) Potential of immunogold-silver staining for the study of leukocyte subpopulations as defined by monoclonal antibodies. J Histochem Cytochem 34:1257–1263

    Google Scholar 

  • De Waele M, De Mey J, Renmans W, Labeur C, Reynaert Ph, Van Camp B (1986) An immunogold-silver staining method for the detection of cell surface antigens in light microscopy. J Histochem Cytochem 34:935–939

    Google Scholar 

  • De Waele M, Renmans W, Segers E, De Valck V, Jochmans K (1989) An immunogold-silver staining method for the detection of cell surface antigens in cell smears. J Histochem Cytochem 37:1855–1862

    Google Scholar 

  • De Waele M, Renmans W, Segers E, Jochmans K, Van Camp B (1988) Sensitive detection of immunogold-silver staining with darkfield and epipolarization microscopy. J Histochem Cytochem 36:679–683

    Google Scholar 

  • Duhamel RC, Johnson DA (1985) Use of nonfat dry milk to block nonspecific nuclear and membrane staining by avidin conjugates. J Histochem Cytochem 33:711–714

    Google Scholar 

  • Ellis IO, Bell J, Bancroft JD (1988) An investigation of optimal gold particle size for immunocytochemical immunogold and immunogold-silver staining to be viewed by polarized indirect light (epipolarization) microscopy. J Histochem Cytochem 36:121–124

    Google Scholar 

  • Faulk WP, Taylor GM (1971) An immunocolloid method for electron microscope. Immunochemistry 8:1081–1083

    Google Scholar 

  • Geoghegan WD, Scillian JJ, Ackerman GA (1978) The detection of human B-lymphocytes by both light and electron microscopy utilizing colloidal gold-labelled anti-immunoglobulin. Immunol Commun 7:1–12

    Google Scholar 

  • Holgate CS, Jackson P, Cowen PN, Bird CC (1983) Immunogoldsilver staining: a new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 31:938–944

    Google Scholar 

  • Horisberger M (1981) Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. In: Johari O (ed) Scanning electron microscopy, vol. 2. SEM Inc. O'Hare, Chicago, p 9

    Google Scholar 

  • Horisberger M, Rosset J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25:295–305

    Google Scholar 

  • Knapp W, Dörken B, Rieber P, Schmidt R, Stein H, Borne A von dem (1989) CD antigens 1989. Blood 74:1448–1450

    Google Scholar 

  • Lah JJ, Hayes DM, Burry RW (1990) A neutral pH silver development method for the visualisation of 1-nanometer gold particles in preembedding electron microscopic immunocytochemistry. J Histochem Cytochem 38:503–508

    Google Scholar 

  • Manara G, Ferari C, Torresani C, Sansoni P, De Panfilis G (1990) The immunogold-silver staining approach in the study of lymphocyte subpopulations in transmission electron microscopy. J Immunol Methods 128:59–63

    Google Scholar 

  • Matutes E, Catovsky D (1982) The fine structure of normal lymphocyte subpopulations — a study with monoclonal antibodies and the immunogold technique. Clin Exp Immunol 50:416–425

    Google Scholar 

  • Pallesen G, Plesner T (1987) The Third International Workshop and Conference on Human Leukocyte Differentiation Antigens with an up-to-date overview of the CD Nomenclature. Leukemia 1:231–234

    Google Scholar 

  • Romasko F, Rosenberg J, Wybran J (1985) An immunogold silver staining method for the light microscopic analyses of blood lymphocyte subsets with monoclonal antibodies. Am J Clin Pathol 84:307–316

    Google Scholar 

  • Romano EL, Romano M (1977) Staphylococcus protein A bound to colloidal gold: a useful reagent to label antigen-antibody sites in electron microscopy. Immunochemistry 14:711–715

    Google Scholar 

  • Roth J (1982) Applications of immunocolloids in light microscopy. J Histochem Cytochem 30:691–696

    Google Scholar 

  • Shaw S (1987) Characterization of human leukocyte differentiation antigens. Immunol Today 8:1–3

    Google Scholar 

  • Vogt RF Jr, Philips DL, Henderson LO, Whitfield W, Spierto FW (1987) Quantitative differences among various proteins as blocking agents for ELISA microtiter plates. J Immunol Methods 101:43–50

    Google Scholar 

  • Wybran J, Rosenberg J, Romasco F (1985) Immunogold-staining: an alternative method for lymphocyte subset enumeration. Comparison with immunofluorescence microscopy and flow cytometry. J Immunol Methods 76:229–238

    Google Scholar 

  • Yokota S (1988) Effect of particle size on labeling density for catalase in protein A-gold immunocytochemistry. J Histochem Cytochem 36:107–109

    Google Scholar 

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De Valck, V., Renmans, W., Segers, E. et al. Light microscopical detection of leukocyte cell surface antigens with a one-nanometer gold probe. Histochemistry 95, 483–490 (1991). https://doi.org/10.1007/BF00315744

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