[3dem] Vitrobot Mark 4 issues
Bob Grassucci
rg2502 at columbia.edu
Thu Feb 19 14:15:33 PST 2009
Hi All,
We just got our FEI Vitrobot Mark IV and I have been trying to get the
parameters worked out for reproduceably making good frozen hydrated
grids. We had a previous version Mark 2 before we moved tho Columbia
and were able to optimize it but the Mark IV has been modified so that
these parameters are no longer valid. We are using Quantifoil grids
with thin carbon spread over them and the ice is either too thick or the
carbon is damaged by plunging. The parameters offset have been changed
to blot force which varies the distance the pads are from each other
when blotting. I have tried blotting times from 1 to 8 seconds and blot
forces from -25 to +20. FEI suggests that I use their blot papers only
but when I tried it there was no improvement. We still are able to make
grids the old fashioned way with our home made device but would like to
take advantage of this expensive apparatus. If anyone else has a
similar set up i.e. Mark IV and using thin carbon, I would appreciate
hearing your experience with it and if you got it to work what
parameters you used. Thanks for your help
Regards,
Bob
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