Department of Materials, Department of Bioengineering and Institute of Biomedical Engineering, Imperial College London, London, SW7 2AZ, UK.
Department of Medicine, Imperial College London, London, W12 0NN, UK.
Adv Mater. 2019 Aug;31(32):e1900488. doi: 10.1002/adma.201900488. Epub 2019 Jun 13.
Volumetric imaging techniques capable of correlating structural and functional information with nanoscale resolution are necessary to broaden the insight into cellular processes within complex biological systems. The recent emergence of focused ion beam scanning electron microscopy (FIB-SEM) has provided unparalleled insight through the volumetric investigation of ultrastructure; however, it does not provide biomolecular information at equivalent resolution. Here, immunogold FIB-SEM, which combines antigen labeling with in situ FIB-SEM imaging, is developed in order to spatially map ultrastructural and biomolecular information simultaneously. This method is applied to investigate two different cell-material systems: the localization of histone epigenetic modifications in neural stem cells cultured on microstructured substrates and the distribution of nuclear pore complexes in myoblasts differentiated on a soft hydrogel surface. Immunogold FIB-SEM offers the potential for broad applicability to correlate structure and function with nanoscale resolution when addressing questions across cell biology, biomaterials, and regenerative medicine.
体积成像技术能够以纳米级分辨率关联结构和功能信息，对于深入了解复杂生物系统中的细胞过程至关重要。最近，聚焦离子束扫描电子显微镜（FIB-SEM）的出现通过对超微结构的体积研究提供了无与伦比的洞察力；然而，它无法以同等分辨率提供生物分子信息。在这里，开发了免疫金 FIB-SEM，它将抗原标记与原位 FIB-SEM 成像相结合，以便能够同时空间映射超微结构和生物分子信息。该方法应用于研究两个不同的细胞材料系统：在微结构基底上培养的神经干细胞中组蛋白表观遗传修饰的定位，以及在软水凝胶表面上分化的成肌细胞中核孔复合物的分布。免疫金 FIB-SEM 具有广泛的适用性，可在解决细胞生物学、生物材料和再生医学领域的问题时，以纳米级分辨率关联结构和功能。
