A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids
- Brookhaven National Lab. (BNL), Upton, NY (United States); Binghamton Univ., NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); North Babylon High School, Babylon, NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); Northport High School, Northport, NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); St Augustine Catholic High School, Markham, ON (Canada)
- Brookhaven National Lab. (BNL), Upton, NY (United States); Earl L. Vandermeulen High School, Port Jefferson, NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); John P. Stevens High School, Edison, NJ (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); Connetquot High School, Bohemia, NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States); PEC Univ. of Technology, Chandigarh (India)
- Brookhaven National Lab. (BNL), Upton, NY (United States)
- Brookhaven National Lab. (BNL), Upton, NY (United States). Photon Sciences Directorate; Brookhaven National Lab. (BNL), Upton, NY (United States). Biosciences Dept.
- Brookhaven National Lab. (BNL), Upton, NY (United States). Photon Sciences Directorate
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 can be instrumental for improving few-nanolitre crystallizations.
- Research Organization:
- Brookhaven National Laboratory (BNL), Upton, NY (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER)
- Grant/Contract Number:
- SC00112704; 11-008; P41RR012408; P41GM103473
- OSTI ID:
- 1228894
- Report Number(s):
- BNL-110969-2015-JA; ACSFEN
- Journal Information:
- Acta Crystallographica. Section F, Structural Biology Communications, Vol. 70, Issue 12; ISSN 2053-230X
- Publisher:
- International Union of CrystallographyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
Using sound pulses to solve the crystal-harvesting bottleneck
|
journal | October 2018 |
Similar Records
High throughput screening using acoustic droplet ejection to combine protein crystals and chemical libraries on crystallization plates at high density
Hitting the target: fragment screening with acoustic in situ co-crystallization of proteins plus fragment libraries on pin-mounted data-collection micromeshes