Federal State Budgetary Institution of Science "M.M. Krasnov Research Institute of Eye Diseases" (Krasnov Research Institute of Eye Diseases), 119021 Moscow, Russia.
Institute of Fundamental Medicine and Biology, Kazan Federal University, 420008 Kazan, Russia.
Int J Mol Sci. 2024 Oct 15;25(20):11076. doi: 10.3390/ijms252011076.
A comprehensive understanding of intercellular and cell-matrix interactions is essential for advancing our knowledge of cell biology. Existing techniques, such as fluorescence microscopy and electron microscopy, face limitations in resolution and sample preparation. Supravital lanthanoid staining provides new opportunities for detailed visualization of cellular metabolism and intercellular interactions. This study aims to describe the structure, elemental chemical, and probable origin of zones of extreme lanthanoid (neodymium) accumulation that form during preparation for scanning electron microscopy (SEM) analysis in corneal fibroblasts filopodia. The results identified three morphological patterns of neodymium staining in fibroblast filopodia, each exhibiting asymmetric staining within a thin, sharp, and extremely bright barrier zone, located perpendicular to the filopodia axis. Semi-quantitative chemical analyses showed neodymium-labeled non-linear phosphorus distribution within filopodia, potentially indicating varying phosphate anion concentrations and extreme phosphate accumulation at a physical or physicochemical barrier. Phosphorus zones labeled with neodymium did not correspond to mitochondrial clusters. During apoptosis, the number of filopodia with extreme and asymmetric phosphorus accumulation increases. Supravital lanthanoid staining coupled with SEM allows detailed visualization of intercellular and cell-matrix interactions with high contrast and resolution. These results enhance our understanding of phosphate anion accumulation and transfer mechanisms in cells under normal conditions and during apoptosis.
全面了解细胞间和细胞基质的相互作用对于推进我们对细胞生物学的认识至关重要。现有的技术，如荧光显微镜和电子显微镜，在分辨率和样品制备方面存在局限性。活体镧系元素染色为详细观察细胞代谢和细胞间相互作用提供了新的机会。本研究旨在描述在角膜成纤维细胞丝状伪足进行扫描电子显微镜（SEM）分析准备过程中形成的极度镧系（钕）积累区域的结构、元素化学性质和可能的起源。结果在成纤维细胞丝状伪足中发现了三种形态的钕染色模式，每个模式在垂直于丝状伪足轴的薄、锐利且极其明亮的屏障区域内呈现不对称染色。半定量化学分析显示丝状伪足内标记有钕的非线性磷分布，可能表明磷酸盐阴离子浓度不同，在物理或物理化学屏障处有极端的磷酸盐积累。用钕标记的磷区与线粒体团簇不对应。在细胞凋亡过程中，具有极度和不对称磷积累的丝状伪足数量增加。活体镧系元素染色与 SEM 相结合，可实现具有高对比度和分辨率的详细细胞间和细胞基质相互作用的可视化。这些结果增强了我们对细胞在正常条件下和细胞凋亡过程中磷酸盐阴离子积累和转移机制的理解。
