[3dem] clip ring in cryo-holder

Kenneth Goldie k.goldie at unibas.ch
Wed Feb 22 09:07:11 PST 2017


Dear Alex

we have a number of 626 holders and they are used daily. On some occasions for screening we may swap out up to 10 grids in a day session per holder so pretty heavy use. With that amount of use from a diverse range of users we find on average we have to replace clip rings or the clip ring holding springs around every 6 months or so, though sometimes a ring may only clip well for 3 months if very heavily used. A holder that is clipped on average twice a day should be fine for over a year form our experience. If a clip ring is not holding a grid tightly you will usually see drift at high magnifications in the EM which may make a sudden circular movement. Normally the rings should really "click" when inserted. Test at room temperature clamping a grid and take a sharp tweezer and touch it on the surface of the grid to see if the grid is held tightly in the holder. If it moves at all, change the clip ring and/or the clip ring spring.  There is a knack to exchanging the clip ring and it is easy to damage your expensive holder so this should only be performed either by Gatan or by someone used to fine mechanical work as the holder recess that holds the clip ring spring may itself get damaged. A jig or stand designed to hold the holder is useful for this job. This is best done under a magnifying head band type loupe or better, a binocular light microscope. If a spring is worn the rounded edges that come into contact with the clip ring will be flattened somewhat, likewise the edges of the clip ring.

Good luck


Ken Goldie, PhD
Facility Manager, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 18| eMail. k.goldie at unibas.ch

Postal Address for delivery of Letters / Goods / Express Carriers:
Ken Goldie | C-CINA, University of Basel, WR0-1058 |
Warenannahme WRO-5.P.057| Mattenstrasse 24a | 4058 Basel, Switzerland

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Today's Topics:

   1. Re: clip ring in cryo-holder (Alexander Shkumatov)
   2. postdoc fellowships in cryo-EM of virus replication
      (Lars-Anders Carlson)
   3. Re: clip ring in cryo-holder (Sharon G  Wolf)


----------------------------------------------------------------------

Message: 1
Date: Wed, 22 Feb 2017 16:55:44 +0100
From: Alexander Shkumatov <ashkumatov at gmail.com>
To: 3dem at ncmir.ucsd.edu
Subject: Re: [3dem] clip ring in cryo-holder
Message-ID: <51B5EBB7-6F71-4183-9C08-EC166AC91171 at gmail.com>
Content-Type: text/plain; charset="utf-8"

sorry, i was not accurate with terms: we see that the spring wire (spring that holds clip ring in place) is kind of getting worn out and somewhat bends which makes it
impossible to insert the clip ring.

We have 626-60. Cornelia, what about clip wire do you have to replace that regularly?

Sharon, what would be the shelf-life of ?springs? (clip wires) from your experience?

Thank you for your answers!
Alex



> On Feb 22, 2017, at 16:16, Sharon G Wolf <Sharon.Wolf at weizmann.ac.il> wrote:
>
> We have both 626-60 and 626-70 holders. The holders are around 15 years old. The rings have NEVER broken, but indeed the springs do need replacing periodically. You can tell that it needs replacing when you see that a brand new clip ring will hold the grid tightly clamped in.  Also one needs to replace the clip rings themselves periodically because otherwise they do not clamp down perfectly and the grids will become contaminated or partially devitrified. We always have extra clip rings ready in stock. This is easier done for the 626-60 than for the 626-70 holders, because the clip rings for the 626-60 are significantly cheaper.
> Its hard for me to imagine the clip rings actually BREAKING. Do they shatter? Do they crack? When does this occur? At what point in the operation?
> Best,
> Sharon
>
>
> From: 3dem [mailto:3dem-bounces at ncmir.ucsd.edu <mailto:3dem-bounces at ncmir.ucsd.edu>] On Behalf Of cornelia at strubi.ox.ac.uk <mailto:cornelia at strubi.ox.ac.uk>
> Sent: Wednesday, 22 February, 2017 15:09
> To: ashkumatov at gmail.com <mailto:ashkumatov at gmail.com>; Reinhard.Rachel at biologie.uni-regensburg.de <mailto:Reinhard.Rachel at biologie.uni-regensburg.de>
> Cc: 3dem at ncmir.ucsd.edu <mailto:3dem at ncmir.ucsd.edu>
> Subject: Re: [3dem] clip ring in cryo-holder
>
> Hi Alex,
>
> Which tilt, 60 or 70 degree holders? They have different cliprings and tools, screw tool (60, thicker clipring, steeper slope) vs 'butterfly' tool (70 degree, thinner ring).
>
> I second more training/supervision, but our 626 70 degree holders are getting a little tired (the spring that holds the clipring in place doesn't seem as firm as it used to. No broken cliprings to date, however). We have, however, ordered new cliprings to see if it helps.
>
> Best wishes,
>
> Cornelia Cazey
>
> ------ Original message------
> From: Alexander Shkumatov
> Date: Wed, 22 Feb 2017 14:57
> To: Reinhard Rachel;
> Cc: 3dem at ncmir.ucsd.edu <mailto:3dem at ncmir.ucsd.edu>;
> Subject:Re: [3dem] clip ring in cryo-holder
>
> Dear Reinhard We are using Gatan 626 cryo-holder and JEOL JEM-1400, respectively. Thank you for a quick reply! Alex> On Feb 22, 2017, at 15:48, Reinhard Rachel  wrote:> >>>> 22.02.2017 <tel:22.02.2017> at 15:43:>> We have a 120kV microscope that is used heavily for screening purposes.> Since >> recently we have about 5-6 active cryo users and >> a clip ring in a cryo holder being broken (not working properly) twice in 3> >> months. I wonder if this is a normal half-life period for clip rings, say, >> it?s expected that it breaks every 2-3 months of heavy usage?> > which clip-ring of specifically which type of cryo-holder in which TEM? this> is critical to know, Alex. > be aware that the new Fischione cryo-holder avoids this type of problem. worth> considering. > kind regards, Reinhard> > (Yes, it sounds that 'users' need some in-depth advice and/or training, or> need to be removed from the user list ...)> > > -- > Prof. Dr. Reinhard Rachel> University of Regensburg> Centre for EM /
  Anatomy> Faculty of Biology & Preclin. Med.> Universitaetsstrasse 31> D-93053 Regensburg - Germany> tel +49 941 943 -2837 <tel:+49%20941%20943%20-2837>, -1720> mail reinhard.rachel at biologie.uni-regensburg.de <mailto:%20reinhard.rachel at biologie.uni-regensburg.de>> office: VKL 3.1.29 > > Next microscopy conferences:> - DGE AK PANOS: Regensburg, 7. April 2017> - Microscopy Conference 2017 (D-A-CH) Lausanne, CH> 20-25 August 2017> http://www.mc2017.ch/ <http://www.mc2017.ch/>> - 19th International Microscopy Congress, Sydney; 9-14 Sept 2018> http://imc19.com/ <http://imc19.com/>> - next Microbiol. conferences: > VAAM/DGHM - Annual Conf > March 5-8, 2017, Wuerzburg> _______________________________________________3dem mailing list3dem at ncmir.ucsd.eduhtt <mailto:%20list3dem at ncmir.ucsd.eduhtt>ps://mail.ncmir.ucsd.edu/mailman/listinfo/3dem <http://mail.ncmir.ucsd.edu/mailman/listinfo/3dem>
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Message: 2
Date: Wed, 22 Feb 2017 17:14:18 +0100
From: Lars-Anders Carlson <lars-anders.carlson at umu.se>
To: <3dem at ncmir.ucsd.edu>
Subject: [3dem] postdoc fellowships in cryo-EM of virus replication
Message-ID: <94944d5c-6fbf-53a1-829e-5a1e1c90f25c at umu.se>
Content-Type: text/plain; charset="utf-8"; format=flowed

Dear all,
Two postdoc fellowships are available in my lab at Ume? University,
Sweden. The projects aim to decipher the molecular basis of human RNA
virus replication using cryo-EM (tomography of cells and single particle
analysis) and in vitro membrane reconstitution.

We are looking for highly motivated candidates with a background in
structural biology, biophysics, biochemistry or a related field. While
prior knowledge of cryo-electron microscopy or in vitro reconstitution
is beneficial, these are the core methods of the lab and we can thus
also provide a postdoc with in situ training specific to the project.

Ume? has just completed the installation of a new EM facility including
a Titan Krios with a phase plate, Falcon III, GIF and K2 Summit, a Scios
FIB/SEM and a Talos 120. Ume? is also an outstanding environment for
molecular infection biology, including the Nordic EMBL Node MIMS and the
Wallenberg Centre for Molecular Medicine.

For more information about the projects and how to apply, got to
http://www.ucmr.umu.se/positions.html

Feel free to contact me directly with any questions!

Best wishes,
Lars


--
Lars-Anders Carlson, Dr. rer. nat.
Assistant Professor
Dept of Medical Biochemistry and Biophysics
Wallenberg Centre for Molecular Medicine
Ume? University

http://www.medchem.umu.se/english/research/principal-investigators/lars-anders-carlson/


------------------------------

Message: 3
Date: Wed, 22 Feb 2017 16:25:29 +0000
From: "Sharon G  Wolf" <Sharon.Wolf at weizmann.ac.il>
To: Alexander Shkumatov <ashkumatov at gmail.com>
Cc: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
Subject: Re: [3dem] clip ring in cryo-holder
Message-ID: <1645E4B0-7DBB-40E9-8FCE-FDE8531C42CC at weizmann.ac.il>
Content-Type: text/plain; charset="windows-1252"

We replace those spring wires once every few years.

Sent from my iPhone

On 22 Feb 2017, at 15:56, Alexander Shkumatov <ashkumatov at gmail.com<mailto:ashkumatov at gmail.com>> wrote:

sorry, i was not accurate with terms: we see that the spring wire (spring that holds clip ring in place) is kind of getting worn out and somewhat bends which makes it
impossible to insert the clip ring.

We have 626-60. Cornelia, what about clip wire do you have to replace that regularly?

Sharon, what would be the shelf-life of ?springs? (clip wires) from your experience?

Thank you for your answers!
Alex



On Feb 22, 2017, at 16:16, Sharon G Wolf <Sharon.Wolf at weizmann.ac.il<mailto:Sharon.Wolf at weizmann.ac.il>> wrote:

We have both 626-60 and 626-70 holders. The holders are around 15 years old. The rings have NEVER broken, but indeed the springs do need replacing periodically. You can tell that it needs replacing when you see that a brand new clip ring will hold the grid tightly clamped in.  Also one needs to replace the clip rings themselves periodically because otherwise they do not clamp down perfectly and the grids will become contaminated or partially devitrified. We always have extra clip rings ready in stock. This is easier done for the 626-60 than for the 626-70 holders, because the clip rings for the 626-60 are significantly cheaper.
Its hard for me to imagine the clip rings actually BREAKING. Do they shatter? Do they crack? When does this occur? At what point in the operation?
Best,
Sharon


From: 3dem [mailto:3dem-bounces at ncmir.ucsd.edu] On Behalf Of cornelia at strubi.ox.ac.uk<mailto:cornelia at strubi.ox.ac.uk>
Sent: Wednesday, 22 February, 2017 15:09
To: ashkumatov at gmail.com<mailto:ashkumatov at gmail.com>; Reinhard.Rachel at biologie.uni-regensburg.de<mailto:Reinhard.Rachel at biologie.uni-regensburg.de>
Cc: 3dem at ncmir.ucsd.edu<mailto:3dem at ncmir.ucsd.edu>
Subject: Re: [3dem] clip ring in cryo-holder

Hi Alex,

Which tilt, 60 or 70 degree holders? They have different cliprings and tools, screw tool (60, thicker clipring, steeper slope) vs 'butterfly' tool (70 degree, thinner ring).

I second more training/supervision, but our 626 70 degree holders are getting a little tired (the spring that holds the clipring in place doesn't seem as firm as it used to. No broken cliprings to date, however). We have, however, ordered new cliprings to see if it helps.

Best wishes,

Cornelia Cazey

------ Original message------
From: Alexander Shkumatov
Date: Wed, 22 Feb 2017 14:57
To: Reinhard Rachel;
Cc: 3dem at ncmir.ucsd.edu<mailto:3dem at ncmir.ucsd.edu>;
Subject:Re: [3dem] clip ring in cryo-holder


Dear Reinhard We are using Gatan 626 cryo-holder and JEOL JEM-1400, respectively. Thank you for a quick reply! Alex> On Feb 22, 2017, at 15:48, Reinhard Rachel  wrote:> >>>> 22.02.2017<tel:22.02.2017> at 15:43:>> We have a 120kV microscope that is used heavily for screening purposes.> Since >> recently we have about 5-6 active cryo users and >> a clip ring in a cryo holder being broken (not working properly) twice in 3> >> months. I wonder if this is a normal half-life period for clip rings, say, >> it?s expected that it breaks every 2-3 months of heavy usage?> > which clip-ring of specifically which type of cryo-holder in which TEM? this> is critical to know, Alex. > be aware that the new Fischione cryo-holder avoids this type of problem. worth> considering. > kind regards, Reinhard> > (Yes, it sounds that 'users' need some in-depth advice and/or training, or> need to be removed from the user list ...)> > > -- > Prof. Dr. Reinhard Rachel> University of Regensburg> Centre for EM / An
 atomy> Faculty of Biology & Preclin. Med.> Universitaetsstrasse 31> D-93053 Regensburg - Germany> tel +49 941 943 -2837<tel:+49%20941%20943%20-2837>, -1720> mail reinhard.rachel at biologie.uni-regensburg.de<mailto:%20reinhard.rachel at biologie.uni-regensburg.de>> office: VKL 3.1.29 > > Next microscopy conferences:> - DGE AK PANOS: Regensburg, 7. April 2017> - Microscopy Conference 2017 (D-A-CH) Lausanne, CH> 20-25 August 2017> http://www.mc2017.ch/> - 19th International Microscopy Congress, Sydney; 9-14 Sept 2018> http://imc19.com/> - next Microbiol. conferences: > VAAM/DGHM - Annual Conf > March 5-8, 2017, Wuerzburg> _______________________________________________3dem mailing list3dem at ncmir.ucsd.eduhtt<mailto:%20list3dem at ncmir.ucsd.eduhtt>ps://mail.ncmir.ucsd.edu/mailman/listinfo/3dem<http://mail.ncmir.ucsd.edu/mailman/listinfo/3dem>

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