Correlative super-resolution fluorescence and metal-replica transmission electron microscopy.
We combine super-resolution localization fluorescence microscopy with transmission electron microscopy of metal replicas to locate proteins on the landscape of the cellular plasma membrane at the nanoscale. We validate robust correlation on the scale of 20 nm by imaging endogenous clathrin (in two and three dimensions) and apply the method to find the previously unknown three-dimensional position of the endocytic protein epsin on clathrin-coated structures at the plasma membrane.
1] National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, Maryland, USA. [2].
Kem A
KA
Sochacki
1] Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA. [2].
Gleb
G
Shtengel
National Institute of Child Health and Human Development, US National Institutes of Health, Bethesda, Maryland, USA.
Schuyler B
SB
van Engelenburg
Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA.
Harald F
HF
Hess
National Heart, Lung, and Blood Institute, US National Institutes of Health, Bethesda, Maryland, USA.
Justin W
JW
Taraska
D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism / Q000648:ultrastructure; D002966:Clathrin / Q000648:ultrastructure; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D043942:Nanotubes / Q000737:chemistry*
D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D008565:Membrane Proteins; C114154:epsin; D006046:Gold