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Title: A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. Ultimately, the results demonstrate that plate lids canmore » be instrumental for improving few-nanolitre crystallizations.« less
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [5] ;  [6] ;  [7] ;  [8] ;  [9] ;  [10] ;  [11]
  1. Brookhaven National Lab. (BNL), Upton, NY (United States); Binghamton Univ., NY (United States)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States); North Babylon High School, Babylon, NY (United States)
  3. Brookhaven National Lab. (BNL), Upton, NY (United States); Northport High School, Northport, NY (United States)
  4. Brookhaven National Lab. (BNL), Upton, NY (United States); St Augustine Catholic High School, Markham, ON (Canada)
  5. Brookhaven National Lab. (BNL), Upton, NY (United States); Earl L. Vandermeulen High School, Port Jefferson, NY (United States)
  6. Brookhaven National Lab. (BNL), Upton, NY (United States); John P. Stevens High School, Edison, NJ (United States)
  7. Brookhaven National Lab. (BNL), Upton, NY (United States); Connetquot High School, Bohemia, NY (United States)
  8. Brookhaven National Lab. (BNL), Upton, NY (United States); PEC Univ. of Technology, Chandigarh (India)
  9. Brookhaven National Lab. (BNL), Upton, NY (United States)
  10. Brookhaven National Lab. (BNL), Upton, NY (United States). Photon Sciences Directorate; Brookhaven National Lab. (BNL), Upton, NY (United States). Biosciences Dept.
  11. Brookhaven National Lab. (BNL), Upton, NY (United States). Photon Sciences Directorate
Publication Date:
OSTI Identifier:
1228894
Report Number(s):
BNL--110969-2015-JA
Journal ID: ISSN 2053-230X; ACSFEN
Grant/Contract Number:
SC00112704; 11-008; P41RR012408; P41GM103473
Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section F, Structural Biology Communications
Additional Journal Information:
Journal Volume: 70; Journal Issue: 12; Journal ID: ISSN 2053-230X
Publisher:
International Union of Crystallography
Research Org:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY crystallization; dehydration; vapor diffusion; high-throughput screening; acoustic droplet ejection; in situ X-ray data collection