Balercia G, Chen S, Bentivoglio M
Institute of Human Anatomy and Histology, University of Verona, Italy.
J Neurosci Methods. 1992 Oct-Nov;45(1-2):87-98. doi: 10.1016/0165-0270(92)90046-g.
Ultrastructural visualization of non-electron-dense fluorescent retrograde neuronal labeling was attempted by means of photo-oxidation. This procedure was used to convert the fluorescence of neurons labeled by the tracers propidium iodide, rhodamine latex microspheres and fluorogold into a stable diaminobenzidine reaction product. The ultrastructural study revealed an accumulation of electron-dense material in these cells both within lysosomes and scattered in the cytoplasmic matrix. Comparison with several different sets of control samples indicated that this material, on the basis of its amount, electron density and appearance, specifically represents the photoconversion reaction product. The effects of the intensity of the fluorescent labeling and of a prolonged photoconversion on the fine structural features of the reaction product are also described and discussed. The present findings indicate that photoconversion can be effectively applied to ultrastructural study of fluorescent retrogradely labeled neurons. The specificity of the photoconversion reaction product should be tested routinely for each fluorochrome and tissue sample.
尝试通过光氧化对非电子致密荧光逆行神经元标记进行超微结构可视化。该方法用于将由示踪剂碘化丙啶、罗丹明乳胶微球和荧光金标记的神经元荧光转化为稳定的二氨基联苯胺反应产物。超微结构研究显示，这些细胞内的溶酶体中和分散在细胞质基质中的电子致密物质有所积累。与几组不同的对照样品比较表明，基于其数量、电子密度和外观，这种物质具体代表光转化反应产物。还描述并讨论了荧光标记强度和延长光转化对反应产物精细结构特征的影响。目前的研究结果表明，光转化可有效应用于荧光逆行标记神经元的超微结构研究。对于每种荧光染料和组织样品，应常规测试光转化反应产物的特异性。
