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.
Bibliographical noteFunding Information:
We thank the following labs at the NIH for technical help with our early trials of cell imaging in PDMS wells. The Morgan lab (NIBIB) for help with design and printing of the PDMS wells; The Samelson lab (NCI) for confocal imaging; The NIAID center for microscopy for STED imaging; The Schwartzberg lab (NHGRI) for providing mouse cells; The Shroff lab (NIBIB) for SMLM imaging. We thank Valarie Barr (NCI) for her edits and comments on the manuscript. This research was supported by Grant no. 321993 from the Marie Skłodowska-Curie actions of the European Commission, the Lejwa Fund, and Grants no. 1417/13, 1937/13, and 1761/17 from the Israeli Science Foundation.
© 2021, The Author(s).