Imaging of Unstained DNA Origami Triangles with Electron Microscopy

Londono‐Calderon, Alejandra; Hossen, Md Mir; Palo, Pierre E.; Bendickson, Lee; Vergara, Sandra; Nilsen‐Hamilton, Marit; Hillier, Andrew C.; Prozorov, Tanya

doi:10.1002/smtd.201900393

Imaging of Unstained DNA Origami Triangles with Electron Microscopy

Journal Article · Tue Aug 13 04:00:00 EDT 2019 · Small Methods

DOI:https://doi.org/10.1002/smtd.201900393· OSTI ID:1557787

Londono‐Calderon, Alejandra ^[1]; Hossen, Md Mir ^[2]; Palo, Pierre E. ^[2]; Bendickson, Lee ^[2]; Vergara, Sandra ^[3]; Nilsen‐Hamilton, Marit ^[2]; Hillier, Andrew C. ^[2]; ^[1]

Ames Lab., Ames, IA (United States)
Ames Lab., Ames, IA (United States); Iowa State Univ., Ames, IA (United States)
Univ. of Pittsburgh, PA (United States)

Imaging of scaffolded DNA and DNA origami nanostructures has been dominated by atomic force microscopy of samples immobilized on bulk substrates. Less commonly used are electron microscopy techniques, typically carried out after negative staining of DNA structures or by using cryo-transmission electron microscopy (TEM). In this work, direct imaging of unstained DNA origami on common electron-transparent substrates with utilizing high angular annular dark field scanning transmission electron microscopy (HAADF-STEM) is reported. This approach establishes a method for depositing and imaging intact DNA triangles with mass-thickness contrast sufficient to measure the scaffold-to-scaffold distances and the length of the triangle's seam. The signal-to-noise ratio (SNR) of the DNA supported on amorphous carbon as a function of the carbon thickness is measured on three types of commercially available TEM grids. This allows for edge detection of ≈1 nm height DNA triangles on carbon substrates as thick as ≈25 nm. Observations on the effect on SNR with the imaging modes are discussed. The effect of cation concentration used for pretreating the grid on the image resolution is also explored. Finally, this work presents proof-of-concept results demonstrating that electron microscopy can be used to resolve key elements of the DNA origami triangle without the use of staining protocols.

View Accepted Manuscript (DOE)

Research Organization:: Ames Laboratory (AMES), Ames, IA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Materials Sciences & Engineering Division

Grant/Contract Number:: AC02-07CH11358

OSTI ID:: 1557787

Alternate ID(s):: OSTI ID: 1557006

Report Number(s):: IS-J--9972

Journal Information:: Small Methods, Journal Name: Small Methods Journal Issue: 12 Vol. 3; ISSN 2366-9608

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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