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A simple technique for immunoelectronmicroscopic localization of Ig on tissue sections with special reference to cutaneous pathology

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Summary

A simple technique using immunoperoxidase-labelled antibodies is reported for the ultrastructural localization of fixed immunoglobulins in tissue lesions. This technique allows the immunolabeling of frozen sections of 15 µ mounted on glass slides. These sections are then closed over for ultrastructural examination, using an Epon-filled gelatin capsule, and then returned to the glass slide. After polymerization, the capsule containing the tissue section is separated from the glass by immersion in liquid nitrogen. This technique is applied to the study of fixed Ig in the skin lesions of lupus erythematosus, pemphigus, bullous pemphigoid and dermatitis herpetiformis. The results obtained show a fine localization of fixed Ig, good tissue conservation and easy orientation of the biological material. It can be applied to the study of tissue antigens.

Zusammenfassung

Eine einfache Technik, die sich der immunperoxydasemarkierten Antikörper bedient, wird für die ultrastrukturelle Lokalisation der Immunoglobuline in Gewebeherden verwandt. Diese Technik erlaubt die Immunmarkierung von Gefrierschnitten von 15 µ Dicke auf Glasträger aufgezogen. Die Schnitte werden dann für die ultrastrukturelle Untersuchung unter Verwendung von Epon-gefüllten gelierten Kapseln überzogen und dann wieder auf den Objektträger gebracht. Nach Polymerisation wird die Kapsel, welche den Gewebeherd enthält, vom Glas durch Eintauchen in flüssigen Stickstoff getrennt. Diese Technik wurde verwandt, um gebundene Antikörper in den Hautherden des Lupus erythematodes, Pemphigus, bullöses Pemphigoid und Dermatitis herpetiformis zu untersuchen. Die Ergebnisse zeigen eine gute Möglichkeit zur Lokalisation der fixierten Antikörper, gute Konservierung des Gewebes und leichte Orientierung des biologischen Materials. Die Methode kann zum Studium von Gewebsantigenen verwandt werden.

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Additional information

Chargé de recherche INSERM. This study was supported by U.E.R. Biologie Humaine, Lyon Offprint requests to: D. Schmitt

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Schmitt, D., Germain, D. & Thivolet, J. A simple technique for immunoelectronmicroscopic localization of Ig on tissue sections with special reference to cutaneous pathology. Arch. Derm. Res. 259, 235–245 (1977). https://doi.org/10.1007/BF00561451

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Key words

  • Immunoelectronmicroscopy
  • Skin
  • SLE
  • Pemphigus
  • Bullous pemphigoid
  • Dermatitis herpetiformis