Hi Jai, I agree with what Pamela says here and want to reiterate that sometimes if you mount a needle longer than the loop itself, the outside part of the needle can cause the part of the needle inside the loop to bend upon freezing, which can destroy or distort the lattice. Best to use a loop bigger than the needle, or just break it into pieces. Acupuncture needles are good for doing this. Good luck! Nicola From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of Pamela J Focia Sent: Monday, September 26, 2016 7:42 PM To: [log in to unmask] Subject: Re: [ccp4bb] Problem in crystals Hello Jai, I worked with long needle crystals for many years. I found that it was perfectly okay if they broke, sometimes much better to use a part of a crystal than to try to harvest/loop them whole. I even sometimes broke them on purpose! When you loop long needle crystals whole they are susceptible to being affected by forces and strain of the cryosolvent and an imperfectly flat loop, or, if you accidentally get part of the crystal on the fiber, it could bow the crystal. Even when I was able to loop an intact crystal, I found that there were better parts of most of the crystals w.r.t. diffraction limit and quality, and usually had to search for the 'sweet spot' to get the best data from each crystal. If you have not done so yet, I recommend trying part of a broken crystal in the beam and see what you get. You might be pleasantly surprised. Good luck! -pamela On Sep 26, 2016, at 8:16 AM, Thomas, Leonard M. <[log in to unmask]<mailto:[log in to unmask]>> wrote: As I tell all students and those new to mounting crystals. It just takes patience and practice. The litho loops from either Mitegen or Molecular Dimensions work pretty well also. Leonard M. Thomas Ph.D. Macromolecular Crystallography Laboratory Oklahoma COBRE in Structural Biology Price Family Foundation Institute of Structural Biology Department of Chemistry and Biochemistry University of Oklahoma Stephenson Life Sciences Research Center 101 Stephenson Parkway Norman, OK 73019 405-325-1126 [log in to unmask]<mailto:[log in to unmask]> https://urldefense.proofpoint.com/v2/url?u=http-3A__barlywine.chem.ou.edu&d=CwIFAg&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=KaQsb7Eep6rBKq4kVaJGW9wZ0byweUXIhZVijB0mDhI&m=1N7gZN2A7aJUWiq2d5xOMqZ_AesLbYeA_xsecWE_qKs&s=0zDwkooKF0vZcDPN8g8C6exUAGGoAkhpg6GflSrliXM&e= https://urldefense.proofpoint.com/v2/url?u=http-3A__structuralbiology.ou.edu&d=CwIFAg&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=KaQsb7Eep6rBKq4kVaJGW9wZ0byweUXIhZVijB0mDhI&m=1N7gZN2A7aJUWiq2d5xOMqZ_AesLbYeA_xsecWE_qKs&s=_MRbvTOUuhz08kDtMnug6YFKF4JPBiV3kli_NhSvsS0&e= On Sep 26, 2016, at 7:25 AM, Bonsor, Daniel <[log in to unmask]<mailto:[log in to unmask]>> wrote: I have had similar crystals which I could harvest using Mitegen Loops http://www.mitegen.com/technotes/handling_needle_crystals.shtml<https://emea01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.mitegen.com%2Ftechnotes%2Fhandling_needle_crystals.shtml&data=01%7C01%7CNicola.1.evans%40KCL.AC.UK%7C338c2c5b21fd4b034b9308d3e63ce36b%7C8370cf1416f34c16b83c724071654356%7C0&sdata=wdXHOHUVyHZm0Bf9HbthI9qjvs3mj2PTvkVQPSi2MCc%3D&reserved=0> FYI I don't work or own shares in Mitegen. Dan Get Outlook for Android<https://emea01.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.proofpoint.com%2Fv2%2Furl%3Fu%3Dhttps-3A__aka.ms_ghei36%26d%3DCwMFAg%26c%3DyHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws%26r%3DKaQsb7Eep6rBKq4kVaJGW9wZ0byweUXIhZVijB0mDhI%26m%3DpBSixOtgLvHkeSl8QQpPrmmsa5ovbgW5CCDoWfBuKVg%26s%3Dwzm6gBZoBQg_lYNiekgJCMTP3SkLsP_oPvYwv8ZfyKk%26e%3D&data=01%7C01%7CNicola.1.evans%40KCL.AC.UK%7C338c2c5b21fd4b034b9308d3e63ce36b%7C8370cf1416f34c16b83c724071654356%7C0&sdata=FGNS%2B55ZMmAkxKLf8MfRZBdRFAngGCt39%2BTsTbG8q6E%3D&reserved=0> ________________________________ From: CCP4 bulletin board <[log in to unmask]<mailto:[log in to unmask]>> on behalf of jai mohan <[log in to unmask]<mailto:[log in to unmask]>> Sent: Monday, September 26, 2016 8:12:20 AM To: [log in to unmask]<mailto:[log in to unmask]> Subject: [ccp4bb] Problem in crystals Dear all, Recently I crystallized a Green Fluorescent Protein (GFP) containing mutations; from the condition PEG8k (w/v), HEPES & MgCl2 or NaCl | protein conc 21mg/ml | crystal growth time 4-6 days. The crystals seems to be thin and breaks immediately during loop out from drop. Hence, I could not go ahead for diffraction, could anyone suggest some inputs. With best regards S.M.Jaimohan. Ph. D <IMG_20160920_150036.jpg> ------- ---- Pamela J. Focia, Ph.D. Research Assistant Professor Structural Biology Facility Manager Robert H. Lurie Comprehensive Cancer Center in the Departments of: Biochemistry and Molecular Genetics, Feinberg School of Medicine, and Molecular Biosciences, Northwestern University 303 E. Chicago Ave., S-215, Chicago 60611 Tarry 7-710, 7-721 c (312)286-3274 o (312)503-0848 f (312)503-5349 [log in to unmask]<mailto:[log in to unmask]>