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

Journal Article · · Acta Crystallographica. Section F, Structural Biology Communications
 [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
+ Show Author Affiliations

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
Citation Metrics:
Cited by: 10 works
Citation information provided by
Web of Science

References (20)

A Semiautomatic Protein Crystallization System with Preventing Evaporation of Drops and Surface Sensor of Solution journal December 2003
Control of the rate of evaporation in protein crystallization by the `microbatch under oil' method journal August 2008
A Linear Relationship between Crystal Size and Fragment Binding Time Observed Crystallographically: Implications for Fragment Library Screening Using Acoustic Droplet Ejection journal July 2014
Real-space refinement of the structure of hen egg-white lysozyme journal January 1974
Coot model-building tools for molecular graphics journal November 2004
Progress in macromolecular crystallography depends on further miniaturization of crystallization experiments journal October 2014
X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection journal May 2011
Crystallization screening: the influence of history on current practice journal June 2014
Lessons from high-throughput protein crystallization screening: 10 years of practical experience journal March 2011
In-plate protein crystallization, in situ ligand soaking and X-ray diffraction journal August 2011
Crystallization of membrane proteins in Cubo book January 2002
Experimental determination of the radiation dose limit for cryocooled protein crystals journal March 2006
Small-Scale Batch Crystallization of Proteins Revisited journal February 2002
Acoustically Mounted Microcrystals Yield High-Resolution X-ray Structures journal May 2011
Liquid–liquid diffusion crystallization improves the X-ray diffraction of EndoS, an endo-β- N -acetylglucosaminidase from Streptococcus pyogenes with activity on human IgG journal November 2013
A procedure for setting up high-throughput nanolitre crystallization experiments. I. Protocol design and validation journal March 2003
Overview of the CCP 4 suite and current developments journal March 2011
Hitting the target: fragment screening with acoustic in situ co-crystallization of proteins plus fragment libraries on pin-mounted data-collection micromeshes journal April 2014
A Droplet-Based, Composite PDMS/Glass Capillary Microfluidic System for Evaluating Protein Crystallization Conditions by Microbatch and Vapor-Diffusion Methods with On-Chip X-Ray Diffraction journal May 2004
Nanoliter-Scale Protein Crystallization and Screening with a Microfluidic Droplet Robot journal May 2014

Cited By (2)

Gentle, fast and effective crystal soaking by acoustic dispensing journal March 2017
Using sound pulses to solve the crystal-harvesting bottleneck journal October 2018

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Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
crystallization
dehydration
vapor diffusion
high-throughput screening
acoustic droplet ejection
in situ X-ray data collection