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Hi Olga,
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<div class="">Should point out there are two versions of this currently out there- the EM GP and EM GP 2. We have the ‘original’ EM GP, ours had no problems reaching 4 °C (would get there in 15-20 mins from RT), but as Denis has rightly pointed this then causes
lots of problems with icing up and the arm not moving properly after a short period of time. We run ours at 8 °C as standard and its fine like this. The EMGP 2 (we have had on trial and I’ve taught on course using it) doesn’t seem to suffer from this so much
and has a nice drainage port at the back which helps a little to drain away any water that does accumulate (although isn’t perfect, still seems to accumulate). </div>
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<div class="">With the humidity, we have also seen this, Denis’s trick of unscrewing the water tank has worked well on occasions for us, but once the humidity sensor needed replacing so if it continues worth a call to Leica! Can be worth sticking a probe in
the side to independently measure the humidity, this will give you an indication of what is really going on. </div>
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<div class="">We have both the Leica EM GP and Vitrobot Mv IV. For single particles in our hands both are capable of performing equivalently, with some tweaking, so I don’t say one is much better than the other for single particles (and users are quite evenly
spread in our facility), but the Leica does have some useful features that can help with some projects;</div>
<div class="">1) You can choose front blotting (filter paper contacts meniscus directly) or back blotting (sample drawn through the grid), which can give you some useful options if you want to concentrate large samples such as filaments of bacterial cells onto
the grid. But for most single particle projects we do front blotting. If you are growing eukaryotic cells onto grids the back blotting option is perfect. </div>
<div class="">2) I’ve found the sensor totally useless on the EM GP, it seems a little better on the GP2 but we still seemed to get more reproducible results without using it. </div>
<div class="">3) Whatever blotting modality you are using, I recommend setting the arm so it makes full, firm and flat contact with the grid, but without bending/flexing the grid at all. We typically use blotting times of 3-5 seconds for single particles using
Whatman 1 paper. I also like to position the tweezer arm more towards the centre of the filter paper circle, this helps to iron out some of the irreproducibility caused when the filter paper ‘ruffles’ after a short period in the humid atmosphere. </div>
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<div class="">Hope this helps, and happy to provide some more info if it would be useful :) Good luck!</div>
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<div class="">Best wishes,</div>
<div class="">Becky</div>
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Rebecca Thompson<br class="">
Senior cryo-Electron Microscopy Support Scientist/Facility manager <br class="">
Astbury Biostructure Laboratory<br class="">
University of Leeds<br class="">
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Email<span class="Apple-tab-span" style="white-space: pre;"> </span><a href="mailto:r.f.thompson@leeds.ac.uk" class="">r.f.thompson@leeds.ac.uk</a><br class="">
Phone<span class="Apple-tab-span" style="white-space: pre;"> </span>0113 3438957/3438959 (office/Titan Krios control room)</div>
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Twitter<span class="Apple-tab-span" style="white-space: pre;"> </span> Bex_16</div>
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<div class="">On 19 Jun 2019, at 20:55, Denis Chrétien <<a href="mailto:denis.chretien@univ-rennes1.fr" class="">denis.chretien@univ-rennes1.fr</a>> wrote:</div>
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The one I have in my possession now did not cool down to 4C for nearly 3 hrs,</div>
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<div class="">We have now been using the EM GP for several years. At 4 °C, we faced problems of ice cristallisation blocking the trap and arm systems. It was « solved » by Leica engineers by increasing the lower temperature that the device can reach (I don’t
remember, between 5 and 7 °C). This setting can be modified in the service menu if you have the code. One way to overcome this problem if it happens is to increase the temperature for a short time so that the ice melts and then come back to 4 °C (after removing
water…). Don’t let the water accumulate and remove it periodically.</div>
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<div class="">and the humidity was going up and down.</div>
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<div class="">Same problem from time to time … One way we overcome this problem is to unscrew the water tank so that air can circulate more freely inside the device. </div>
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<div class="">Is it the normal behaviour? </div>
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<div class="">I won’t say « normal », but yes it can behaves like that :-(</div>
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<div class="">Also, what is the good regime for protein solutions blotting? Could it be compared to Vitrobot? </div>
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<div class="">No comparison, I never used a Vitrobot. Nevertheless, what’s very cool with the Leica is the temperature control of the liquid ethane. We set it to -184 °C and it can last hours at this temperature without refilling.</div>
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<div class="">Hope that helps, waiting also for the feedback of other users,</div>
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<div class="">Thank you!</div>
<div class="">Olga</div>
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=======<br class="">
Yours sincerely,<br class="">
Olga Sokolova, PhD, D.Sc.<br class="">
Professor RAS, <span class="" style="font-size: 12.8px;">Faculty of biology </span><span class="" style="font-size: 12.8px;">Moscow State University</span></div>
<div dir="ltr" class="">phone +7-495-938-0005<br class="">
cell +7-916-4841251<br class="">
<a href="http://proteom-msu.org/en/index.html" target="_blank" class="">http://proteom-msu.org/en/index.html<br class="">
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