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Title: Challenges and solutions for the analysis of in situ , in crystallo micro-spectrophotometric data

Combining macromolecular crystallography with in crystallo micro-spectrophotometry yields valuable complementary information on the sample, including the redox states of metal cofactors, the identification of bound ligands and the onset and strength of undesired photochemistry, also known as radiation damage. However, the analysis and processing of the resulting data differs significantly from the approaches used for solution spectrophotometric data. The varying size and shape of the sample, together with the suboptimal sample environment, the lack of proper reference signals and the general influence of the X-ray beam on the sample have to be considered and carefully corrected for. In the present article, we discuss how to characterize and treat these sample-dependent artefacts in a reproducible manner and we demonstrate the SLS-APE in situ, in crystallo optical spectroscopy data-analysis toolbox.
 [1] ;  [2] ;  [3] ;  [4]
  1. Paul Scherrer Inst. (PSI), Villigen (Switzerland). Swiss Light Source
  2. Univ. of Essex, Colchester (United Kingdom)
  3. European Molecular Biology Lab. (EMBL), Hamburg (Germany)
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Photon Sciences Directorate
Publication Date:
Report Number(s):
Journal ID: ISSN 1399-0047; ABCRE6
Grant/Contract Number:
SC00112704; 20111166; 2370
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section D: Biological Crystallography (Online)
Additional Journal Information:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography (Online); Journal Volume: 71; Journal Issue: 1; Journal ID: ISSN 1399-0047
International Union of Crystallography
Research Org:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
47 OTHER INSTRUMENTATION; micro-spectrophotometer; UV-visible absorption; Raman spectroscopy; single-crystal spectroscopy; software toolbox; data analysis; background correction
OSTI Identifier: