Size perspective of nanodroplets

Published in reference J. Nowakowski, C.N. Cronin, D.E. McRee, M.W. Knuth, C.G. Nelson, N.P. Pavletich, J. Rogers, B.-C. Sang, D.N. Scheibe, R.V. Swanson, D.A. Thomspon "Structures of the cancer-related Aurora-A, FAK, and EphA2 protein kinases from nanovolume crystallography " Structure, 10, 1659-1667 (2002).

Images of the Nanovolume Crystallization Droplets and Microcrystals Used for Protein Structure Determination
(A) Fragment of the crystallization plate showing the size of a 50 nl (white arrow) and the actual crystals of (B) FAK, (C) Aurora-A, and (D) EphA2 used to collect the X-ray diffraction data. d_min indicates the resolution of the data collected from each crystal.

Crystallization using various drop sizes

Published in reference B.D. Santarsiero, D.T. Yegian, C.C. Lee, G. Spraggon, J. Gu, D. Scheibe, D.C. Uber, E.W. Cornell, R.A. Nordmeyer, W.F. Kolbe, J. Jin, A.L. Jones, J.M. Jaklevic, P. G. Schultz, R.C. Stevens "An approach to rapid protein crystallization using nanodroplets" J. Appl. Crystallography, 35, 278-281 (2002).

Representative images of lysozyme crystallization trials, obtained at 4ºC, with various total drop volumes. The protein solution and reservoir solution drop volumes used to form the crystallization drops are equivalent: (A) 2 μL drop volume, 48-72 hours for initial crystallization; (B) 1 μL, 36-48 h; (C) 500 nL, 24-36 h; (D) 200 nL, 18 h; (E) 40 nL, 10-12 h; (F) 20 nL, 2 h. (G) magnified view of (F) showing crystal formation, roughly 50 microns on the longest edge.

Published in reference D. Hosfield, J. Palan, M. Hilgers, D. Scheibe, D. McRee, R.C. Stevens. "A Fully Integrated Protein Crystallization Platform for Small Molecule Drug Discovery" J. Struct. Biol. 142, 207-217, (2003).

Time course of crystal growth. A series of images of the same crystal drop over 11 days is presented. The drop consists of 50 nl protein solution
mixed with 50 nl of well solution. A small crystal starts to grow in the lower part of the drop on day 1 and has grown much larger on day 3. On day 5 the crystal has started to pit and by day 11 the crystal has shrunk considerably and is pitted. Noting this degradation, the next time this protein is set up, the plate can be harvested on day 3 and the crystal frozen to preserve it from further degradation.

Robots for Cloning

Robots for Fermentation

4th generation 96-well fermentor (above)	1st generation 96-well fermentor (above)
1st generation 96-well baculovirus fermentor (above)

Robots for Purification

2nd generation protein purification (above)	1st generation protein purification (above)

Robots for Crystallization

2nd generation crystallization robot (Agincourt - above)	1st generation crystallization robot (Agincourt - above)

Robots for Imaging

Protein crystal incubator and imager Fort (above)	1st generation imaging system (above)

last updated December 19, 2003 - send corrections and comments to Angela Walker (alwalker@scripps.edu)