title: Automated harvesting and processing of protein crystals through laser photoablation.
abstract: Currently, macromolecular crystallography projects often require the use of highly automated facilities for crystallization and X-ray data collection. However, crystal harvesting and processing largely depend on manual operations. Here, a series of new methods are presented based on the use of a low X-ray-background film as a crystallization support and a photoablation laser that enable the automation of major operations required for the preparation of crystals for X-ray diffraction experiments. In this approach, the controlled removal of the mother liquor before crystal mounting simplifies the cryocooling process, in many cases eliminating the use of cryoprotectant agents, while crystal-soaking experiments are performed through diffusion, precluding the need for repeated sample-recovery and transfer operations. Moreover, the high-precision laser enables new mounting strategies that are not accessible through other methods. This approach bridges an important gap in automation and can contribute to expanding the capabilities of modern macromolecular crystallography facilities.
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zander
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Hoffmann
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Irina
authors.initials: I
authors.lastname: Cornaciu
authors.affiliation: Structure Design Informatics and Structural Biology, Sanofi, 13 Quai Jules Guesde, 94403 Vitry-sur-Seine, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Marquette
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Gergely
authors.initials: G
authors.lastname: Papp
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Landret
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Gaël
authors.initials: G
authors.lastname: Seroul
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Jérémy
authors.initials: J
authors.lastname: Sinoir
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Martin
authors.initials: M
authors.lastname: Röwer
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Frank
authors.initials: F
authors.lastname: Felisaz
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Sonia
authors.initials: S
authors.lastname: Rodriguez-Puente
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Mariaule
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Peter
authors.initials: P
authors.lastname: Murphy
authors.affiliation: Structure Design Informatics and Structural Biology, Sanofi, 13 Quai Jules Guesde, 94403 Vitry-sur-Seine, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Mathieu
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Cipriani
authors.affiliation: Grenoble Outstation, European Molecular Biology Laboratory; Unit of Virus Host-Cell Interactions (UMI 3265), University Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, 38042 Grenoble, France.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Márquez
mesh_terms: D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods*; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D045647:DNA Glycosylases / Q000737:chemistry*; D006801:Humans; D007834:Lasers
chemical_list: D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human
keywords: automated cryocooling; automated crystal mounting; crystal soaking; ligand screening