Dear all,
I would like to know how to break down protein crystal agglomerates. My crystallisation experiments yield quite a high number of crystal agglomerates (see images attached for 10x and 20x magnification). So far I am sonicating my sample for ~13 mins to break down crystal agglomerates but it does not seem to work (see attached image before_sonification for before and after comparison). I wonder whether you folks have any idea of how to separate the crystals to obtain images of single crystals (If you would suggest surfactants, which surfactants would you suggest?). I would need single/ individual crystals as I am developing a MATLAb® routine to derive the crystal size distribution from imaging crystals with an optical microscope.
With thanks and kindest regards,
Frederik
Dear Frederik,
Long sonication times (more than a few minutes) might break the crystals or even promote secondary nucleation (this might explain why you observe more agglomerates after sonication). I would try pulsed sonication instead.
However, I would first focus on the crystallisation conditions and crystallisation volume.
Good luck,
Joana
along with Joanas recommendation, I would reduce protein concentration in front of crystallization. Further you might try to to settle out larger agglomerates. Sonication is certainly not the answer to your problem.
Frederik Link
Use glycerol to stabilize the dispersion of the protein crystal structure
Article Use of glycerol, polyols and other protein structure stabili...
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