Biophysics, Center for Integrative Physiology and Molecular Medicine (CIPMM), School of Medicine, Saarland University, Homburg, Germany.
INM-Leibniz Institute for New Materials, Saarbrücken, Germany.
Front Immunol. 2022 Jan 28;13:828634. doi: 10.3389/fimmu.2022.828634. eCollection 2022.
Visualizing interactions between cells and the extracellular matrix (ECM) mesh is important to understand cell behavior and regulatory mechanisms by the extracellular environment. However, long term visualization of three-dimensional (3D) matrix structures remains challenging mainly due to photobleaching or blind spots perpendicular to the imaging plane. Here, we combine label-free light-sheet scattering microcopy (LSSM) and fluorescence microscopy to solve these problems. We verified that LSSM can reliably visualize structures of collagen matrices from different origin including bovine, human and rat tail. The quality and intensity of collagen structure images acquired by LSSM did not decline with time. LSSM offers abundant wavelength choice to visualize matrix structures, maximizing combination possibilities with fluorescently-labelled cells, allowing visualizing of long-term ECM-cell interactions in 3D. Interestingly, we observed ultrathin thread-like structures between cells and matrix using LSSM, which were not observed by normal fluorescence microscopy. Transient local alignment of matrix by cell-applied forces can be observed. In summary, LSSM provides a powerful and robust approach to investigate the complex interplay between cells and ECM.
可视化细胞与细胞外基质 (ECM) 网格之间的相互作用对于理解细胞行为和细胞外环境的调节机制非常重要。然而，由于光漂白或垂直于成像平面的盲点，长期可视化三维 (3D) 基质结构仍然具有挑战性。在这里，我们结合无标记光片散射显微镜 (LSSM) 和荧光显微镜来解决这些问题。我们验证了 LSSM 可以可靠地可视化来自不同来源的胶原基质结构，包括牛、人、鼠尾胶原。LSSM 获取的胶原结构图像的质量和强度不会随时间下降。LSSM 提供了丰富的波长选择来可视化基质结构，与荧光标记细胞的组合可能性最大化，允许在 3D 中可视化长期的 ECM-细胞相互作用。有趣的是，我们使用 LSSM 观察到细胞和基质之间的超细线状结构，而普通荧光显微镜则观察不到这些结构。可以观察到细胞施加的力引起的基质的瞬时局部排列。总之，LSSM 为研究细胞与 ECM 之间的复杂相互作用提供了一种强大而稳健的方法。
