Racah Institute of Physics, The Hebrew University, Jerusalem, Israel.
Graduate Program in Biophysics, Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Department of Physics, Harvard University, Cambridge, MA, USA.
Commun Biol. 2021 Apr 1;4(1):439. doi: 10.1038/s42003-021-01960-2.
Cell-cell interfaces convey mechanical and chemical information in multicellular systems. Microscopy has revealed intricate structure of such interfaces, yet typically with limited resolution due to diffraction and unfavourable orthogonal orientation of the interface to the coverslip. We present a simple and robust way to align cell-cell interfaces in parallel to the coverslip by adhering the interacting cells to two opposing coverslips. We demonstrate high-quality diffraction-limited and super-resolution imaging of interfaces (immune-synapses) between fixed and live CD8 T-cells and either antigen presenting cells or melanoma cells. Imaging methods include bright-field, confocal, STED, dSTORM, SOFI, SRRF and large-scale tiled images. The low background, lack of aberrations and enhanced spatial stability of our method relative to existing cell-trapping techniques allow use of these methods. We expect that the simplicity and wide-compatibility of our approach will allow its wide dissemination for super-resolving the intricate structure and molecular organization in a variety of cell-cell interfaces.
细胞-细胞界面在多细胞系统中传递机械和化学信息。显微镜已经揭示了这种界面的复杂结构，但由于衍射和界面与盖玻片之间不利的正交方向，通常分辨率有限。我们提出了一种简单而强大的方法，通过将相互作用的细胞粘附到两个相对的盖玻片上，使细胞-细胞界面与盖玻片平行排列。我们展示了高质量的衍射极限和超分辨率成像，用于固定和活的 CD8 T 细胞与抗原呈递细胞或黑色素瘤细胞之间的界面（免疫突触）。成像方法包括明场、共聚焦、STED、dSTORM、SOFI、SRRF 和大规模平铺图像。与现有的细胞捕获技术相比，我们的方法具有低背景、无像差和增强的空间稳定性，允许使用这些方法。我们预计，我们方法的简单性和广泛的兼容性将使其广泛传播，用于解析各种细胞-细胞界面中复杂的结构和分子组织。
