title: Radiation damage in protein crystals is reduced with a micron-sized X-ray beam.
abstract: Radiation damage is a major limitation in crystallography of biological macromolecules, even for cryocooled samples, and is particularly acute in microdiffraction. For the X-ray energies most commonly used for protein crystallography at synchrotron sources, photoelectrons are the predominant source of radiation damage. If the beam size is small relative to the photoelectron path length, then the photoelectron may escape the beam footprint, resulting in less damage in the illuminated volume. Thus, it may be possible to exploit this phenomenon to reduce radiation-induced damage during data measurement for techniques such as diffraction, spectroscopy, and imaging that use X-rays to probe both crystalline and noncrystalline biological samples. In a systematic and direct experimental demonstration of reduced radiation damage in protein crystals with small beams, damage was measured as a function of micron-sized X-ray beams of decreasing dimensions. The damage rate normalized for dose was reduced by a factor of three from the largest (15.6 μm) to the smallest (0.84 μm) X-ray beam used. Radiation-induced damage to protein crystals was also mapped parallel and perpendicular to the polarization direction of an incident 1-μm X-ray beam. Damage was greatest at the beam center and decreased monotonically to zero at a distance of about 4 μm, establishing the range of photoelectrons. The observed damage is less anisotropic than photoelectron emission probability, consistent with photoelectron trajectory simulations. These experimental results provide the basis for data collection protocols to mitigate with micron-sized X-ray beams the effects of radiation damage.
authors: National Institute of General Medical Sciences and National Cancer Institute Collaborative Access Team, Biosciences Division, Argonne National Laboratory, Argonne, IL 60439, USA.
authors: Ruslan
authors: R
authors: Sanishvili
authors: Derek W
authors: DW
authors: Yoder
authors: Sudhir Babu
authors: SB
authors: Pothineni
authors: Gerd
authors: G
authors: Rosenbaum
authors: Shenglan
authors: S
authors: Xu
authors: Stefan
authors: S
authors: Vogt
authors: Sergey
authors: S
authors: Stepanov
authors: Oleg A
authors: OA
authors: Makarov
authors: Stephen
authors: S
authors: Corcoran
authors: Richard
authors: R
authors: Benn
authors: Venugopalan
authors: V
authors: Nagarajan
authors: Janet L
authors: JL
authors: Smith
authors: Robert F
authors: RF
authors: Fischetti
mesh_terms: D016880:Anisotropy; D018360:Crystallography, X-Ray / Q000706:statistics & numerical data; D009010:Monte Carlo Method; D011506:Proteins / Q000737:chemistry* / Q000528:radiation effects*
chemical_list: D011506:Proteins