Müller T
Institute for Anatomy, University of Mainz, Germany.
Biotech Histochem. 1994 May;69(3):171-6. doi: 10.3109/10520299409106281.
Methylene blue and neutral red were selected for staining mast cell granules by supravital injections. A new technique was applied for embedding in paraffin and Araldite without dislocation or loss of dye. Stabilization and electron microscopic identification of the dyes were achieved by transforming them into electron-dense precipitates using phosphomolybdic acid dissolved in a paraformaldehyde-glutaraldehyde mixture to preserve the ultrastructure of the tissues. It was found that in general the intensity of the light microscopic staining correlated directly with the electron density. Closer study revealed that not all cytoplasmic granules exhibited the same strong affinity for the cationic dyes. Furthermore, differences in dye distribution were observed within the granules themselves. The difference in the staining pattern can be explained by the heterogeneous occurrence of the anionic residues. Because of its high sensitivity and relatively low toxicity, the method described here is well suited for detecting the binding sites of organic cations in tissues under supravital or vital conditions.
通过活体注射选择亚甲蓝和中性红对肥大细胞颗粒进行染色。应用了一种新的石蜡和环氧树脂包埋技术，染料无错位或损失。通过将染料溶解在多聚甲醛 - 戊二醛混合物中的磷钼酸转化为电子致密沉淀物来实现染料的稳定化和电子显微镜鉴定，以保存组织的超微结构。发现一般来说，光学显微镜染色的强度与电子密度直接相关。进一步研究表明，并非所有细胞质颗粒对阳离子染料都表现出相同的强亲和力。此外，在颗粒内部观察到染料分布的差异。染色模式的差异可以通过阴离子残基的异质存在来解释。由于其高灵敏度和相对低毒性，这里描述的方法非常适合在活体或生命条件下检测组织中有机阳离子的结合位点。
