Whole-cell imaging of plasma membrane receptors by 3D lattice light-sheet dSTORM

Felix Wäldchen,Michael Luciano,Jan Schlegel,Sören Doose,Markus Sauer,Ralph Götz,Martin Schnermann

doi:10.1038/s41467-020-14731-0

Abstract

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.

Highlights

The molecular organization of receptors in the plasma membrane of cells is paramount for their functionality
We used lattice light-sheet (LLS) microscopy[9,10,11,12] in combination with single-molecule localization microscopy (SMLM), by direct stochastic optical reconstruction microscopy[13] or photoactivated localization microscopy (PALM)[14] and single-particle tracking for whole-cell 3D imaging of plasma membrane receptors[15,16]
Our data disclose that localization densities are in general lower at the basal plasma membrane, and CD56 and CD45 accumulate at cell–cell interfaces

Summary

Introduction

The molecular organization of receptors in the plasma membrane of cells is paramount for their functionality. We used lattice light-sheet (LLS) microscopy[9,10,11,12] in combination with single-molecule localization microscopy (SMLM), by direct stochastic optical reconstruction microscopy (dSTORM)[13] or photoactivated localization microscopy (PALM)[14] and single-particle tracking for whole-cell 3D imaging of plasma membrane receptors[15,16].

Results

Conclusion