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Modern structural biology relies on NMR, X-ray crystallography andcryo-electron microscopy for gaining information on biomolecules at nanometer,sub-nanometer or atomic resolution. All these methods, however, requireaveraging over a vast ensemble of entities and hence knowledge on theconformational landscape of an individual particle is lost. Unfortunately,there are now strong indications that even X-ray free electron lasers will notbe able to image individual molecules but will require nanocrystal samples.Here, we show that non-destructive structural biology of single particles hasnow become possible by means of low-energy electron holography. As an example,individual tobacco mosaic virions deposited on ultraclean freestanding grapheneare imaged at one nanometer resolution revealing structural details arisingfrom the helical arrangement of the outer protein shell of the virus. Sincelow-energy electron holography is a lens-less technique and since electronswith a deBroglie wavelength of approximately 1 Angstrom do not impose radiationdamage to biomolecules, the method has the potential for Angstrom resolutionimaging of single biomolecules.

Low-energy electron holographic imaging of individual tobacco mosaic virions