title: A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids.
abstract: 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. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations.
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Zipper
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Xavier
authors.initials: X
authors.lastname: Aristide
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Dylan P
authors.initials: DP
authors.lastname: Bishop
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Ishita
authors.initials: I
authors.lastname: Joshi
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Kharzeev
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Krishna B
authors.initials: KB
authors.lastname: Patel
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Brianna M
authors.initials: BM
authors.lastname: Santiago
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Karan
authors.initials: K
authors.lastname: Joshi
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Kahille
authors.initials: K
authors.lastname: Dorsinvil
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Sweet
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
authors.forename: Alexei S
authors.initials: AS
authors.lastname: Soares
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D011506:Proteins / Q000737:chemistry
chemical_list: D011506:Proteins
keywords: acoustic droplet ejection; crystallization; dehydration; high-throughput screening; in situ X-ray data collection; vapor diffusion