Data from: SUB-immunogold-SEM reveals nanoscale distribution of submembranous epitopes

Electron microscopy paired with immunogold labeling is the most precise
tool for protein localization. However, these methods are either
cumbersome, resulting in small sample numbers and restricted
quantification, or limited to identifying protein epitopes external to the
membrane. Here, we introduce SUB-immunogold-SEM, a scanning electron
microscopy technique that detects intracellular protein epitopes proximal
to the membrane. We identified four critical sample preparation factors
that contribute to the method’s sensitivity and validated its efficacy
through precise localization and high-powered quantification of
cytoskeletal and transmembrane proteins. We evaluate the capabilities of
SUB-immunogold-SEM on cells with highly differentiated apical surfaces:
(i) auditory hair cells, revealing the presence of nanoscale MYO15A-L
rings at the tip of stereocilia; and (ii) respiratory multiciliate cells,
mapping the distribution of the SARS-CoV-2 receptor ACE2 along the motile
cilia. SUB-immunogold-SEM extends the application of SEM-based nanoscale
protein localization for the first time to intracellular epitopes on the
exposed surfaces of any cell.