Circular DNA molecules imaged in air by scanning force microscopy.
Routine and reproducible imaging of DNA molecules in air with the scanning force microscope (SFM) has been accomplished. Circular molecules of plasmid DNA were deposited onto red mica and imaged under various relative humidities. In related experiments, the first images of the Escherichia coli RNA polymerase-DNA complex have also been obtained. This has been possible by (1) the use of specially modified SFM tips with a consistent radius of curvature of 10 nm or less, to minimize the amount of image distortion introduced by the finite dimensions of commercially available tips, (2) the optimization of a method to deposit and bind DNA molecules to the mica surface in a stable fashion, and (3) careful control of the sample humidity, to prevent solvation of the molecules and detachment from the surface by the scanning tip or stylus. Contact forces in the range of a few nanonewtons are routinely possible in air and in the presence of residual humidity. The spatial resolution of the images appears determined by the radius of curvature of the modified styli, which can be estimated directly from the apparent widths of the DNA molecules in the images.
Bustamante
C
C
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Vesenka
J
J
Tang
C L
CL
Rees
W
W
Guthold
M
M
Keller
R
R
D000388:Air; D004262:DNA Restriction Enzymes; D004270:DNA, Circular / Q000378:metabolism / Q000648:ultrastructure*; D004268:DNA-Binding Proteins / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism / Q000648:ultrastructure; D004926:Escherichia coli / Q000201:enzymology; D006813:Humidity; D008855:Microscopy, Electron, Scanning / Q000379:methods; D010957:Plasmids; D015203:Reproducibility of Results
D004270:DNA, Circular; D004268:DNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes