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Figure 4
Automated crystal mounting and cryocooling. (ad) A series of images taken along the time course of the automated harvesting process are shown. (1) The crystallization drop before harvesting; the location of the pin (round oval), the aperture (U-shapes, green) and the cutting area have been selected. (2) The aspiration aperture has been created and the crystallization solution removed by aspiration. (3) The pin has been applied to the film and the laser excision has been performed; the samples are ready for transfer to the cryojet. (4) Cryocooled samples as seen from the camera at the cryojet position. As can be appreciated, crystals tend to remain in position throughout the mounting process. This makes it possible to perform sequential harvesting operations from a single drop (a) or to mount multiple crystals on a single pin (b, c). (e) Crystal surgery: crystals of the Vps34 lipid kinase in complex with compound 3 consistently grow as tight needle-like clusters (top panel). The laser was used to cut and mount a single crystal fragment (the time course is shown from top to bottom). The full process is shown in Supplementary Movie S3. X-ray helical data collection from this sample on the ESRF ID23-2 microfocus beamline produced a high-resolution structure. The bottom panel shows a region of the electron-density OMIT map contoured at 1σ around the compound 3 binding site. Complete data-collection statistics are described in Table 2[link].

Journal logoSTRUCTURAL
ISSN: 2059-7983
Volume 72| Part 4| April 2016| Pages 454-466
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