*** For details on how to be removed from this list visit the ***
*** CCP4 home page http://www.ccp4.ac.uk ***
Rebecca,
I think the success of mounting out of microbatch depends on the type of
oil and plate you are using. We successfully are able to transfer crystals
out of drops and into cryoprotectants using a standard hampton mounted
cryoloop. You need a microscope that has a decent distance between the
plate and actual lens apparatus so you can get the cryoloop down far
enough into the well without covering up your viewpoint. When the drop
contains the cryoprotectant already... then we directly transfer to liquid
nitrogen and do not try and remove the oil.... at that point the oil only
covers the solution which surrounds your crystal. One of the main benifits
of vapour diffusion under oil vs. other sitting drop type methods is that
the crystal (at least in our arrangement/apparatus) never sticks to the
bottom, since the drop is "spericilized" by the oil and barely touches the
plastic. Go to
http://www.bioc.aecom.yu.edu/labs/blanlab/VETTING/PROTOCOLS/vapdiffunderoil.html
if you want to check out our VDUO setup.
Matthew Vetting
On Tue, 12 Sep 2006, Page, Rebecca wrote:
*** For details on how to be removed from this list visit the ***
*** CCP4 home page http://www.ccp4.ac.uk ***
CCP4 community:
I find mounting crystals grown in microbatch drops difficult compared to
mounting crystals grown out of sitting drops, primarily because of the oil
layer. Do most people simply use the oil layer as a cryoprotectant? If not, do
you typically try to remove the oil prior to transferring the crystal to a
cryoprotectant. I'd like to compile 'tricks of the trade' for mounting crystals
out of microbatch plates. Any advice would be helpful and I'll compile the list
of responses and repost for the community.
Thanks in advance,
Rebecca
