title: Automated screening of 2D crystallization trials using transmission electron microscopy: a high-throughput tool-chain for sample preparation and microscopic analysis.
abstract: We have built and extensively tested a tool-chain to prepare and screen two-dimensional crystals of membrane proteins by transmission electron microscopy (TEM) at room temperature. This automated process is an extension of a new procedure described recently that allows membrane protein 2D crystallization in parallel (Iacovache et al., 2010). The system includes a gantry robot that transfers and prepares the crystalline solutions on grids suitable for TEM analysis and an entirely automated microscope that can analyze 96 grids at once without human interference. The operation of the system at the user level is solely controlled within the MATLAB environment: the commands to perform sample handling (loading/unloading in the microscope), microscope steering (magnification, focus, image acquisition, etc.) as well as automatic crystal detection have been implemented. Different types of thin samples can efficiently be screened provided that the particular detection algorithm is adapted to the specific task. Hence, operating time can be shared between multiple users. This is a major step towards the integration of transmission electron microscopy into a high throughput work-flow.
authors.affiliation: MIPS Laboratory, University of Haute Alsace, Mulhouse, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Coudray
authors.forename: Gilles
authors.initials: G
authors.lastname: Hermann
authors.forename: Daniel
authors.initials: D
authors.lastname: Caujolle-Bert
authors.forename: Argyro
authors.initials: A
authors.lastname: Karathanou
authors.forename: Françoise
authors.initials: F
authors.lastname: Erne-Brand
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Buessler
authors.forename: Pamela
authors.initials: P
authors.lastname: Daum
authors.forename: Juergen M
authors.initials: JM
authors.lastname: Plitzko
authors.forename: Mohamed
authors.initials: M
authors.lastname: Chami
authors.forename: Urs
authors.initials: U
authors.lastname: Mueller
authors.forename: Hubert
authors.initials: H
authors.lastname: Kihl
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Urban
authors.forename: Andreas
authors.initials: A
authors.lastname: Engel
authors.forename: Hervé-W
authors.initials: HW
authors.lastname: Rémigy
mesh_terms: D003460:Crystallization / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000379:methods*
chemical_list: D008565:Membrane Proteins