Use of two different methods for the electron microscope radioautographic detection of labeled antibodies bound to thin sections.

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Abstract
Thyroglobulin antigenicity was located by exposure to labeled antibodies [Ab] of thin sections of rat thyroids fixed in glutaraldehyde and embedded in glycol methacrylate. Two methods were used for the handling of sections during Ab exposure and emulsion coating. In the 1st method, sections were labeled while floating freely within plastic "O" rings over solutions of 125I-labeled anti-thyroglobulin Ab and were then mounted on celloidin coated slides to be processed for EM radioautography by the dipping technique. In the 2nd method, sections were first mounted on EM grids that were then floated over the 125I-labeled Ab solution; the sections were then coated with emulsion by means of a wire loop. Qualitative and quantitative comparison of the results obtained with the 2 methods showed that they basically gave the same results, e.g., they disclosed the same pattern of specific labeling within the follicular colloid, colloid droplets, apical vesicles, Golgi and rough endoplasmic reticulum. A slightly lower half distance value for the 2nd method (340 .ANG.) as compared to the first (460 .ANG.) indicated that the 2nd method may provide a slightly better resolution, presumably because only with the 2nd method, the labeled section surface is in immediate contact with the emulsion. Both methods may provide a reasonably satisfactory detection of radioactive antibodies.

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