To Boaz: I seperated the large crystals and checked its diffraction already. While the diffraction is quite poor, only several dots could be seen. T_T I washed the seeds twice with my buffer and seed the drop immediately after setting it. Thanks for your advices and I will try the additives.
To Charles, Enrico, Bernie & Tiantian: Thanks for your kindly advices, I set different conditions for the buffers with glycerol and different protein/reservoir volume ratios already following your instructions. :) To David: Hmm...my crystals are smaller than the smallest loop T_T. It's quite hard for me to pick them up (due to my clumsy fingers...lol). Thanks for your advices and the review. I have another question: I usually stored my protein samples aliquots at -80 degree and thaw the small aliquots when I need to use. While my senior said it will harm the protein so she suggested to keep them at 4 degree. So it is possible that I got the small crystals coz the freezing and thawing alter the proteins? Thanks very much. SnowDeer On Tue, Sep 6, 2011 at 9:39 PM, David Waterman <dgwater...@gmail.com> wrote: > Dear SnowDeer, > > Just how small is too small? If you have access to a microfocus beamline > you might find you can collect decent diffraction data from crystals with > dimensions in the single digit microns. Fishing tiny crystals is difficult, > but something like the MicroMesh "tennis racquet" mounts can help. Having > multiple crystals on the same pin is in fact a rather helpful way of > screening lots of samples. Please don't discount your crystals _only_ > because they are small! > > Shameless plug: there is a review on microcrystallography here that you > might find interesting. > http://www.tandfonline.com/doi/abs/10.1080/0889311X.2010.527964 > > Best wishes > > -- David > > > > On 5 September 2011 09:06, SnowDeer <huanxu...@gmail.com> wrote: > >> Dear All: >> >> Recently I am working on a protein which can already grow nice >> pyramid-like crystals after the condition was optimized, while the crystals >> are too small to be picked up. The crystals grew quite fast and densely, so >> I tried to put 100ul paraffin oil inside the 600ul reservoir solution or put >> the plate under 16 degree to slow down the evaporation, while the crystals >> were still the same. I also tried macro or micro seeding with or without the >> paraffin oil. Macroseeding would give a larger crystal (not very nice) with >> many small crystals in the drop even I washed the seeds carefully. For >> microseeding, the same small crystals grew. >> >> I don't have many experiences in crystallography, so I have no idea how to >> make it grow bigger... >> >> Any suggestion is most welcome. >> >> Thanks. >> SnowDeer >> > >
