Title
Optimization Protocol for the SSRL In-Situ Crystallization and Automated Data Collection Plate
Recommended Citation
October 1, 2017.
Abstract
Typically when researchers want to collect data from protein crystals, those crystals must be flash-frozen using liquid nitrogen. This keeps the crystals stable for data analysis, however, can damage crystals and does not give the best possible data. Ideally, crystals can be analyzed at room temperature, which would allow for more modulation of the shape and give researchers a better picture of the shape of the protein. Room temperature data analysis can be difficult, because protein crystals are fragile and will dehydrate quickly if not kept in the proper conditions. The purpose of this research was to develop and optimize a plate that could store crystals at room temperature without damaging the crystals. Over the course of the project many different conditions were tested to try to find an appropriate environment for the protein crystals inside the SSRL In-Situ Crystallization Plate that would allow crystals to remain viable at room temperature for an extended period of time. It was found that if the crystals were loaded into a plate that had been equilibrated with a crystallization solution thickened with agarose, the crystals would remain viable for at least three days. Diffraction patterns were taken from various crystals after one, two, and three days in order to confirm the crystals were indeed viable and that they refracted properly. The end goal of this project is to allow users to SLAC to be able to utilize this plate and the Stanford Automated Mounter located at SSRL to collect data from room temperature crystals remotely without damage to those crystals.
Mentor
Aina Cohen
Lab site
SLAC National Accelerator Laboratory (SLAC)
Funding Acknowledgement
This material is based upon work supported by the National Science Foundation through the Robert Noyce Teacher Scholarship Program under Grant # 1340110. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. The research was also made possible by the California State University STEM Teacher and Researcher Program, in partnership with Chevron (www.chevron.com), the National Marine Sanctuary Foundation (www.marinesanctuary.org) and SLAC National Accelerator Laboratory.
URL: https://digitalcommons.calpoly.edu/star/439