Towards automated screening of two-dimensional crystals.
Screening trials to determine the presence of two-dimensional (2D) protein crystals suitable for three-dimensional structure determination using electron crystallography is a very labor-intensive process. Methods compatible with fully automated screening have been developed for the process of crystal production by dialysis and for producing negatively stained grids of the resulting trials. Further automation via robotic handling of the EM grids, and semi-automated transmission electron microscopic imaging and evaluation of the trial grids is also possible. We, and others, have developed working prototypes for several of these tools and tested and evaluated them in a simple screen of 24 crystallization conditions. While further development of these tools is certainly required for a turn-key system, the goal of fully automated screening appears to be within reach.
Cheng
Anchi
A
The National Resource for Automated Molecular Microscopy, Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, CB-129, La Jolla, CA 92037, USA. acheng@scripps.edu
Leung
Albert
A
Fellmann
Denis
D
Quispe
Joel
J
Suloway
Christian
C
Pulokas
James
J
Abeyrathne
Priyanka D
PD
Lam
Joseph S
JS
Carragher
Bridget
B
Potter
Clinton S
CS
D001331:Automation; D003460:Crystallization; D003461:Crystallography / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation / Q000379:methods*; D016624:Negative Staining / Q000379:methods; D011510:Proteolipids / Q000648:ultrastructure; D012371:Robotics / Q000295:instrumentation*
D008565:Membrane Proteins; D011510:Proteolipids; C020485:proteoliposomes