Hi Morgan Elizabeth,
You could try this protocol
<https://scripts.iucr.org/cgi-bin/paper?S1600576722002382> (paper title
below, shameless plug), it can cryo-protect your crystals without the
need of touching the crystal drop, while at the same time it is likely
to make your crystals more robust and diffracting to higher resolution.
A drug-discovery-oriented non-invasive protocol for protein
crystal cryoprotection by dehydration, with application for
crystallization screening
<https://scripts.iucr.org/cgi-bin/paper?S1600576722002382>
Good luck!
D
On 23/11/2023 09:52, SHEPARD William wrote:
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Hi Morgan,
Cases like yours are the motivation why the PROXIMA 2A beamline
(amongst other beamlines) have developed a plate screener for
collecting X-ray diffraction data in situ. I suggest you contact those
beamlines to find out how to get your plates in the beam.
Cheers,
Bill
------------------------------------------------------------------------
*From: *"Blake, Morgan Elizabeth" <more...@uab.edu>
*To: *"CCP4BB" <CCP4BB@JISCMAIL.AC.UK>
*Sent: *Wednesday, 22 November, 2023 17:44:29
*Subject: *[ccp4bb] Fragile Crystals
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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