April 30 – June 24, 2014
A facility for cryocooling crystals under pressure
is now available at MacCHESS. This technique, developed in the Gruner lab,
was reported in (Kim et al., Acta Cryst. D61, 881 (2005)). It involves
mounting a crystal on a special pin, pressurizing it, cooling to liquid
nitrogen temperature, and then releasing the pressure while keeping the
crystal cold. The method can allow successful cryocooling using little
or no penetrating cryoprotectant, and can produce cryocooled crystals
of better quality than the usual cryocooling method. In addition,
pressure-cryocooling can act to stabilize a single conformation of a bound
ligand, hence making it visible in an electron density map (Albright et al.,
Cell 126, 1147 (2006)). It is also possible to apply the method to samples
in capillaries, both solutions and crystals mounted, or grown, in
An apparatus for cryocooling samples under pressure has been installed at CHESS.
It is designed to enclose all high pressure components in a steel safety container with
1/2 inch thick walls. Weight: ~3000 lb. Users can provide unfrozen crystals and request staff
to pressure-cool them. Diffraction from the first user crystals processed at CHESS improved in resolution
from 3.2 to 2.8 A, and images showed better spot shapes.
Steps in standard pressure-cooling:
Having constructed and tested the necessary equipment at CHESS,
we are now making pressure-cryocooling available to the user community
on an experimental basis. Please read some important
If you have some crystals that freeze poorly, and you would like
to try this new technique, or if you would just like more
Chae Un Kim,
Chae Un Kim, Jennifer L. Wierman, Richard Gillilan, Enju Lima, Sol M. Gruner.
A New Reduced Background Crystal Hydration Method for High Pressure Cryocooling.
J. Appl. Cryst. (2013) 46, 234-241 (pdf)
Chae Un Kim, Mark W. Tate, and Sol M. Gruner. Protein Dynamical Transition at 110 K.
Proc. Natl. Acad. Sci. USA (2011) 108, 20897-20901. (pdf)
Chae Un Kim, Buz Barstow, Mark W. Tate, and Sol M. Gruner. Evidence for liquid water during the high-density to low-density amorphous ice transition.
PNAS (2009), 106, 4596-4600 (.pdf 1.7 MB )
Chae Un Kim, Yi-Fan Chen, Mark W. Tate and Sol M. Gruner. Pressure-induced high-density amorphous ice in protein crystals.
J. Appl. Cryst. (2008), 41, 1-7. (.pdf 280 kb)
Chae Un Kim, Quan Hao and Sol M. Gruner. Solution of protein crystallographic structures by high-pressure cryocooling and noble-gas phasing.
Acta Cryst. (2006). D62, 687–694. doi:10.1107/S0907444906014727 (.pdf 613 kb)
Chae Un Kim, Quan Hao and Sol M. Gruner. High-pressure cryocooling for capillary sample cryoprotection and diffraction phasing at long wavelengths .
Acta Cryst. (2007). D63, 653-659. doi:10.1107/S0907444907011924 (.pdf 900 kb)
Chae Un Kim, Raphael Kapfer and Sol M. Gruner. High-pressure cooling of protein crystals without cryoprotectants.
Acta Cryst, D61, 2005, 881–890. (.pdf 863 kb)
Direct questions, suggestions or problems to