Advances in phase plate cryo-EM imaging of DNA and nucleosomes.
Contrast in electron cryo-microscopy (cryo-EM) is limited by the weak phase and radiation sensitive nature of biologic samples embedded in vitrified ice. We have recently shown that a new contrast enhancement technique utilizing the Volta phase plate can be combined with single particle analysis to determine the structure of a small chromatin complex, the nucleosome core particle, at near-atomic resolution. Here, we discuss advantages and limitations of the technique in terms of data collection, particle detection, and visualization of individual DNA molecules and higher-order chromatin structure.
a School of Biological Sciences , Nanyang Technological University , Singapore.
Eugene Y D
EYD
Chua
a School of Biological Sciences , Nanyang Technological University , Singapore.|b NTU Institute of Structural Biology , Nanyang Technological University , Singapore.
Sara
S
Sandin
D020285:Cryoelectron Microscopy / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*
D009707:Nucleosomes; D004247:DNA
DNA; Volta phase plate; chromatin; cryo-EM; nucleosome