Dear Elizabeth,
/In situ/ data collection is a good approach to try in your case. You
could use the CrystalDirect technology for automated crystal harvesting
that is more gentle to crystals than manual harvesting. This is
available both at EMBL Grenoble and EMBL Hamburg facilities, which offer
integrated crystallography services in collaboration with the ESRF and
Petra III synchrotrons.
doi:10.3791/62491.
doi:10.1016/j.crmeth.2021.100102.
doi:10.1107/s0907444912031459.
Best wishes
Josan
_________________________________________________________
Jose A. Marquez, Senior Scientist
Head of the Crystallization Facility
European Molecular Biology Laboratory, Grenoble.
Delivery address: EMBL, 71, Avenue des Martyrs
38000 Grenoble, France
Postal address: EMBL, 71, Avenue des Martyrs
CS 90181 38042 Grenoble Cedex 9, France
Phone +33 (0)476 20 74 25
Fax. +33 (0)476 20 71 99
https://www.embl.org/groups/marquez/
https://www.embl.org/services-facilities/grenoble/high-throughput-crystallisation/
https://htxlab.embl.org/
_________________________________________________________
On 11/22/2023 5:44 PM, Blake, Morgan Elizabeth wrote:
Hello!
I am a PhD student working on a crystallography project to wrap up my
dissertation research. I have purified a complex of two proteins, and
I can consistently grow crystals in 10% PEG3350, 0.2M KSCN, 0.1M
BIS-TRIS propane pH 7.5. These crystals have sharp edges and can grow
to a large size (greater than 0.5 mm), but the crystals seem to be
very fragile. When we open the drops to harvest the crystals, we have
little time to harvest the crystals before they crack. When we move
the crystals to a cryoprotectant, over time they start fracturing.
We've tried using different percentages of glycerol, ethylene glycol,
PEG400, and oil for cryoprotectants with no success. Needless to say,
the crystals do not diffract well, with spot patterns that look very
streaky/mosaic, which I presume is due to the defects that we see in
harvesting/handling. We have screened for alternate crystallization
conditions, but we seem to get the same morphology in other
conditions. Does anyone have suggestions for additives we could use
post-crystallization to help stabilize our crystals?
Thanks for your advice!
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