[3dem] 626 holder problems

[Garry P Morgan]

Ruben,
you are correct!  it’s apoferritin…
when you saw this, was the contamination there from the very beginning of your session, or did it appear later?  or gradually get worse?  do you remember what the repair entailed?  was it just new o-rings? or was there more to it?
thanks
Garry


On Mar 5, 2018, at 2:43 PM, Diaz-Avalos, Ruben <diazavalosr at janelia.hhmi.org> wrote:

> Hi Garry,
> 
> I have seen something very similar to this before, and at that time, we concluded that it was heavy contamination from the rod assembly that operates the shutter on the holder. I sent the holder to repair and did not see the problem anymore. 
> 
> The sample looks like apoferritin.
> 
> Ruben.
> 
>> On Mar 5, 2018, at 4:22 PM, Garry P Morgan <Garry.Morgan at Colorado.EDU> wrote:
>> 
>> Hello list!
>> 
>> i have yet another round of questions regarding a problem with one of my Gatan 626 holders.  i believe my holder tip is heating up, but i’m getting no indication of a temperature change on my control box.  i have attached a couple images…first i would like to ask if anyone can acknowledge that the second image (named “5hours_in_TEM”) does in fact look like a de-vitrified sample.
>> 
>> the first image is what my sample looked like initially, and after ~3 hours in the TEM; both images are from the same grid square. my nitrogen was topped up every 1-2 hours (so i’m confident that it didn’t run out), and when i noticed the change in the ice, i immediately checked the temperature with the control box and it was still at -184º (which is the usual reading for us).  likewise, there was no unusual amount of drift during this transformation (as one might expect if the holder was running out of LN2). anyway, my main questions is:  does anyone know what might be causing this?  i’m almost certain something is not right in my holder, but i just got it back (for the second time), so i’m fairly confident all the o-rings as well as the clip spring are (or should be) new and in good shape.
>> 
>> one possible clue: when i first put the rod into the TEM, the column vacuum would not recover until i opened the shutter, at which point the vacuum quickly started to improve.  i guess this would indicate that the o-rings on the shutter assembly rod are not sealing completely…could this lead to a very slow/gradual warmup of the sample? later in my session, opening and closing the shutter had no impact on the column vac.
>> 
>> any thoughts or possible solutions would be greatly appreciated.  and just for fun….can anyone guess what this sample is?
>> 
>> thanks,
>> Garry
>> 
>> 
>> <2hours_in_TEM.jpeg><5hours_in_TEM.jpeg>
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