Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD, 21702, USA.
Nat Commun. 2020 Feb 14;11(1):887. doi: 10.1038/s41467-020-14731-0.
The molecular organization of receptors in the plasma membrane of cells is paramount for their functionality. We combined lattice light-sheet (LLS) microscopy with three-dimensional (3D) single-molecule localization microscopy (dSTORM) and single-particle tracking to quantify the expression and distribution, and mobility of CD56 receptors on whole fixed and living cells, finding that CD56 accumulated at cell-cell interfaces. For comparison, we investigated two other receptors, CD2 and CD45, which showed different expression levels and distributions in the plasma membrane. Overall, 3D-LLS-dSTORM enabled imaging and single-particle tracking of plasma membrane receptors with single-molecule sensitivity unperturbed by surface effects. Our results demonstrate that receptor distribution and mobility are largely unaffected by contact to the coverslip but the measured localization densities are in general lower at the basal plasma membrane due to partial limited accessibility for antibodies.
细胞等离子膜受体的分子组织对于其功能至关重要。我们结合晶格层光片（LLS）显微镜与三维（3D）单分子定位显微镜（dSTORM）和单粒子追踪技术，定量测量了整个固定和活细胞上 CD56 受体的表达、分布和迁移率，发现 CD56 在细胞-细胞界面处聚集。为了进行比较，我们还研究了另外两种受体 CD2 和 CD45，它们在等离子膜上的表达水平和分布不同。总的来说，3D-LLS-dSTORM 能够以单分子灵敏度对等离子膜受体进行成像和单粒子追踪，而不会受到表面效应的干扰。我们的结果表明，受体的分布和迁移率在很大程度上不受与盖玻片接触的影响，但由于抗体的部分有限可及性，基底等离子膜上的测量定位密度通常较低。
