In situ strategy for biomedical target localization via nanogold nucleation and secondary growth.
In situ strategy for biomedical target localization via nanogold nucleation and secondary growth.
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DOI：
10.1038/s42003-021-02246-3
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发表时间：
2021-06-10
影响因子：
5.9
通讯作者：
Hishikawa Y
中科院分区：
文献类型：
--
作者：
Sawaguchi A;Kamimura T;Takahashi N;Yamashita A;Asada Y;Imazato H;Aoyama F;Wakui A;Sato T;Choijookhuu N;Hishikawa Y
免疫细胞化学使目标分子的精确空间位置可视化。超微结构免疫细胞化学最常见的策略是将纳米金颗粒与抗体结合作为探针。然而，传统的纳米金标记需要耗时的纳米金探针制备和细胞/组织样品的超薄切片。在这里，我们介绍了一种涉及通用石蜡/低温恒温器切片上免疫酶产品中纳米金成核的原位策略，并为热湿空气条件下纳米金的开发提供了独特的见解。纳米金颗粒专门定位在肾足细胞上以靶向突触蛋白。透射电子显微镜揭示了二次生长和自组装，可以通过牛血清白蛋白稳定和磷酸盐缓冲盐水加速来进行实验控制。胃 H+/K+-ATP 酶的有价值的回顾性纳米金标记是在 15 年后的老式免疫酶沉积物上实现的（即 15 年沉积物）。目前的原位纳米金标记预计将填补光学显微镜和电子显微镜之间的空白，以关联细胞/组织的结构和功能。泽口等人。介绍一种新颖的免疫细胞化学方法，该方法利用通用石蜡/冷冻切片上的免疫酶产品中的原位金纳米颗粒标记。该方法不仅有助于克服传统纳米金标记方法的局限性，而且有助于弥合光学显微镜和电子显微镜之间的差距。
Immunocytochemistry visualizes the exact spatial location of target molecules. The most common strategy for ultrastructural immunocytochemistry is the conjugation of nanogold particles to antibodies as probes. However, conventional nanogold labelling requires time-consuming nanogold probe preparation and ultrathin sectioning of cell/tissue samples. Here, we introduce an in situ strategy involving nanogold nucleation in immunoenzymatic products on universal paraffin/cryostat sections and provide unique insight into nanogold development under hot-humid air conditions. Nanogold particles were specifically localized on kidney podocytes to target synaptopodin. Transmission electron microscopy revealed secondary growth and self-assembly that could be experimentally controlled by bovine serum albumin stabilization and phosphate-buffered saline acceleration. Valuable retrospective nanogold labelling for gastric H+/K+-ATPase was achieved on vintage immunoenzymatic deposits after a long lapse of 15 years (i.e., 15-year-old deposits). The present in situ nanogold labelling is anticipated to fill the gap between light and electron microscopy to correlate cell/tissue structure and function. Sawaguchi et al. introduce a novel immunocytochemistry method that utilizes in situ gold nanoparticle labelling in immunoenzymatic products on universal paraffin/cryostat sections. The method is helpful, not only in overcoming limitations of the conventional nano gold labelling methods, but also useful in bridging the gap between light and electron microscopy.
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