yes ,i had try both small and big crystals ,and they were same situation ,and i freeze in the LN2.does that mean my crystals are not suit in freeze circumstance ,can i try RM diffraction ? 在2016-07-25 08:30:40,张士军[log in to unmask]写道: How big are your crystals? Try small er ones if you used huge ones. 50%MPD freezes clear in my hands. Do you freeze in the stream or LN2? Jürgen ...................... Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-2926 http://lupo.jhsph.edu On Jul 24, 2016, at 8:11 PM, 张士军 <[log in to unmask]> wrote: Hi all I have a crystal grown in 50%MPD 0.2M sodium citrate pH7.2 ,because MPD can be a cyrobuffer ,so I harvested them directly .But the spots are smearing when I difftacted it .then I add 5%glycerol in the cyrobuffer when harvested it ,the result is the same .Can anyoone give me some suggestion about this situation .Thanks a lot !!!!! ShijunZhang School of Life Science Xiamen University